Studies

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View/Edit 1700100004 2017-03-08 CC Kidney Proximal Tubule [Fluidic] Fluidic-3D -No Diseases- 105 0 0 0 0 0 0 Study of 9 chips, 6 for ammoniagenesis with 3 control chips. Three chips dropped pH and 3 held at 7.4. TC2T Admin Sandra Karcher 4 2018-03-22 Cirit_MIT 2
View/Edit 1700200002 2017-03-29 CC Blood-Brain Barrier (NVU) Fluidic-3D -No Diseases- 32 0 0 0 0 0 1 MIT NVU/BBB study with four chips. This study only assayed FITC-Dextran permeability of the BBB TC2T Michael Castiglione 4 2018-01-16 Cirit_MIT 3
View/Edit 1700100008 2017-04-04 CC Kidney Proximal Tubule [Fluidic] Fluidic-3D -No Diseases- 260 0 0 0 0 0 1 Compare the effect of cell density of the BioPredic RPTEC line on standard KPT viability measures KIM-1 and PrestoBlue TC2T Michael Castiglione 4 2018-03-22 Cirit_MIT 4
View/Edit 1700100010 2017-04-04 CC Kidney Proximal Tubule [MIT, Triple Channel] Fluidic-3D -No Diseases- 81 0 0 0 0 0 1 This study characterizes Lonza RPTEC in the Himmelfarb Kidney Proximal Tubule model by observing the baseline levels of PrestoBlue activation and KIM-1 production over eight days. TC2T Michael Castiglione 4 2018-03-22 Cirit_MIT 5
View/Edit 1700200005 2017-04-04 CC Blood-Brain Barrier (NVU) Fluidic-3D -No Diseases- 156 0 0 0 0 0 1 MIT NVU/BBB study with 4 chips with only the vascular endothelial cells. Assay FITC-dextran permeability without any compounds. TC2T Michael Castiglione 4 2018-03-22 Cirit_MIT 6
View/Edit 1700200006 2017-05-01 CC Blood-Brain Barrier (NVU) Fluidic-3D -No Diseases- 160 0 0 0 0 0 1 MIT NVU/BBB study with 4 chips each with endothelial cells, astrocytes, and pericytes. Chips were assayed for FITC-Dextran permeability before and after adding cells. TC2T Michael Castiglione 4 2018-03-22 Cirit_MIT 7
View/Edit 1700100005 2017-04-04 TOX Kidney Proximal Tubule [96-well Static] Static-2D -No Diseases- 231 0 0 0 0 0 2 Toxicity testing of Lonza RPTEC in two 96-well plates with Cadmium Chloride, Cisplatin, Gentamicin, and Rifampicin. Cell viability was assayed by measuring KIM-1 production, and PrestoBlue reagent modifcation. This study was used to find acceptable concentrations of those four compounds for use in the 3D Kidney Promixal Tubule MPS TC2T Michael Castiglione 4 2018-03-22 Cirit_MIT 8
View/Edit 1700100009 2017-06-12 TOX Kidney Proximal Tubule [MIT, Triple Channel] Fluidic-3D -No Diseases- 782 0 0 0 0 0 0 Enter later. Toxicity study of Kidney Proximal Tubule model (HIM-31 cells). Assay the effect on KIM-1 production from various concentrations of cisplatin, gentamicin, cadmium chloride, and rifampicin TC2T Michael Castiglione 4 2018-01-16 Cirit_MIT 9
View/Edit 1700300005 2017-06-14 TOX Brain Static-3D -No Diseases- 612 0 0 0 0 0 1 MIT Toxicity Study of the Thomson Brain MPS. Chips were exposed to Bortezomib (0.1, 0.001, or 0.0001 uM), or Tamoxifen (1, 0.1, or 0.01) for 14 days. TC2T Michael Castiglione 4 2018-03-22 Cirit_MIT 10
View/Edit 1700300006 2017-07-06 TOX Brain Static-3D -No Diseases- 918 0 0 0 0 0 1 A toxicity study of the Thompson Brain MPS with Docetaxel (10, 1, and 0.1 uM), Doxycycline (10, 1, and 0.1 uM), and Vincristine (1, 0.1, and 0.001 uM). The effect of these compounds were study on Park7, LDH, and N-Acetylaspartic Acid production. Cell viability was assayed throughout the study by the PrestoBlue commercial assay kit. Study Results: Invalid study due to lack of tissue differentiation. TC2T Michael Castiglione 4 2018-04-04 Cirit_MIT 11
View/Edit 1700100012 2017-11-19 TOX Kidney Proximal Tubule [MIT, Triple Channel] Fluidic-3D -No Diseases- 750 0 0 0 0 0 1 This study examines the effect of three compounds on the UW Kidney Proximal Tubule Model. The compounds tested were Gentamicin, Rifampicin, Cadmium dichloride, and Cisplatin, with four concentrations for each compound.The model was assayed for cell viability with PrestoBlue, and for cell injury by measuring secreted KIM-1. TC2T Michael Castiglione 4 2018-03-22 Cirit_MIT 12
View/Edit 1700200008 2017-06-07 TOX-CC Blood-Brain Barrier (NVU) Fluidic-3D -No Diseases- 1,520 0 0 0 0 0 1 MIT's eighth run of four NVU/BBB devices. This study looked at permeability of the membrane over 21 days. It also studied the effect of acute (1 day) 100 ug/mL LPS treatment on membrane permeability and on production of 10 standard human cytokines. TC2T Michael Castiglione 4 2018-03-22 Cirit_MIT 13
View/Edit Media Optimization 2015-05-14 CC SQL-SAL 1.0 Fluidic-3D -No Diseases- 144 0 0 0 0 0 0 Testing media and Quantum Dots UPDDI Felipe Lee Montiel 4 2019-07-22 Taylor_MPS 16
View/Edit Nortis-HAR-V 2015-09-16 CC Pitt-MGH Liver Acinus Fluidic-3D -No Diseases- 28 0 0 0 0 0 0 Testing of the Nortis HAR-V device using cryopreserve hepatocytes, TMNK-1, THP-1 and LX 2 cells UPDDI Felipe Lee Montiel 4 2019-07-22 Taylor_MPS 20
View/Edit Motif-PS-Chipshop 2015-09-30 CC Pitt-MGH Liver Acinus Fluidic-3D -No Diseases- 44 0 0 0 0 0 0 Motif polystyrene devices, using different flow setups. Human hepatocytes fresh isolated were used in. (3 cell model) UPDDI Felipe Lee Montiel 4 2019-07-22 Taylor_MPS 23
View/Edit iPSC-Heps-Nortis 2015-10-06 CC SQL-SAL 1.0 Fluidic-3D -No Diseases- 6 0 0 0 0 0 0 Testing iPSC-Heps SQL-SAL development in Nortis HAR-V UPDDI Felipe Lee Montiel 4 2019-07-22 Taylor_MPS 24
View/Edit PS-Cross-flow-configurations 2015-10-27 CC SQL-SAL 1.0 Fluidic-3D -No Diseases- 122 0 0 0 0 0 0 Testing different perfusion configurations on chipshop PS cross flow devices UPDDI Felipe Lee Montiel 4 2019-07-22 Taylor_MPS 29
View/Edit TCTC Reference Study 2018-01-30 CC SQL-SAL 1.5 Fluidic-3D -No Diseases- 329 0 0 0 0 0 0 This study is a reference study done at the UPDDI for comparison with the SQL-SAL 1.5 models run for the TCTC project. Method: Determine chip to chip reproducibility in vehicle treated (1% DMSO) and 40 µM tolcapone treated SQL-Sal 1.5 devices during 14 day incubation. Results: chip to chip variation was at excellent to acceptable levels for albumin, urea and LDH. Comment: previous studies in this model showed significant reduction in liver functions at 88 and 220 uM tolcapone. The lack of the effect at 40 uM should be retested. UPDDI Dillon Gavlock 4 2018-08-30 Taylor_MPS 40
View/Edit Clone 1 MCF7 ESR1 Mutant Growth Initial 2017-05-22 DM LAMPS MCF7 Metastatic Breast Cancer Model Fluidic-3D -No Diseases- 15 0 0 0 0 0 0 MCF7 ESR1 mutant cells [WT-2 mCherry, Y537S-1 mCherry, D538G-1, mCherry] initial studies cells grown in complete media with no estrogen deprivation (+E2); (Flow Rate: 15 uL/hr) UPDDI Dillon Gavlock 4 2019-08-13 Taylor_MPS 52
View/Edit Clone 1 MCF7 ESR1 Mutant Growth 1 2017-08-07 DM LAMPS MCF7 Metastatic Breast Cancer Model Fluidic-3D -No Diseases- 114 0 90 0 0 0 0 MCF7 ESR1 mutant cells [WT-2 mCherry, Y537S-1 mCherry, D538G-1, mCherry] -/+ 5nM estradiol (E2) (Flow Rate: 15 uL/hr) UPDDI Dillon Gavlock 4 2019-08-13 Taylor_MPS 53
View/Edit Clone 1 MCF7 ESR1 Mutant Growth 2 2017-09-15 DM LAMPS MCF7 Metastatic Breast Cancer Model Fluidic-3D -No Diseases- 144 0 90 0 0 0 0 MCF7 ESR1 mutant cells [WT-2 mCherry, Y537S-1 mCherry, D538G-1, mCherry] -/+ 5nM estradiol (E2) (Flow Rate: 15 uL/hr). While both mutants (Y537S and D538G) demonstrate increased constitutive growth compared to WT in the absence of estrogen, the Y537S muatnt shows enhanced estrogen dependent growth compared to the D538G mutant. This is similar to observations made in co-culture models, but different to phenotypes observed in monoculture where the D538G mutant shows estrogen-enhanced growth. UPDDI Dillon Gavlock 4 2019-08-13 Taylor_MPS 57
View/Edit Clone 2 MCF7 ESR1 Mutant Growth 1 2017-10-16 DM LAMPS MCF7 Metastatic Breast Cancer Model Fluidic-3D -No Diseases- 144 0 0 0 0 0 0 examined growth of second independent set of clones of MCF7 ESR1 mutant cell lines. While both mutants (Y537S and D538G) demonstrate increased constitutive growth compared to WT in the absence of estrogen, the Y537S muatnt shows enhanced estrogen dependent growth compared to the D538G mutant. This is similar to observations made in co-culture models, but different to phenotypes observed in monoculture where the D538G mutant shows estrogen-enhanced growth. In addition, these findings replicate the results obtained using the first set of ESR1 mutant clones. UPDDI Dillon Gavlock 4 2019-08-13 Taylor_MPS 58
View/Edit Drug Recovery from Nortis and Chipshop Devices 2015-11-04 CC LAMPS [Cell Free] Fluidic-3D -No Diseases- 11 0 0 0 0 0 0 72 hr continuous infusion, 72 hr washout study in cell free devices UPDDI Larry Vernetti 4 2019-08-13 Taylor_MPS 114
View/Edit Acetaminophen 15 uL/h vs. 5 uL/h 2016-06-22 TOX LAMPS Fluidic-3D -No Diseases- 478 0 0 0 0 0 0 Comparing acetaminophen toxicity at two concentrations and two different flow rates with controls. UPDDI Luke Bergenthal 4 2019-07-16 Taylor_MPS 116
View/Edit EPA 1 2016-08-04 TOX SQL-SAL 1.5 Fluidic-3D -No Diseases- 256 0 0 0 0 0 0 3 week study comparing Tolcapone and Entacapone UPDDI Albert Gough 4 2019-08-06 Taylor_EPA 117
View/Edit EPA 2 2016-09-15 TOX SQL-SAL 1.5 Fluidic-3D -No Diseases- 343 0 0 0 0 0 0 Eighteen day study assessing the the toxicity of Caffeine, Valproic Acid, Thalidomide, and Warfarin. UPDDI Dillon Gavlock 4 2019-07-16 Taylor_EPA 120
View/Edit EPA 3 2016-09-29 TOX SQL-SAL 1.5 Fluidic-3D -No Diseases- 345 0 0 0 0 0 0 Eighteen day study assessing toxicity of Buspirone, Metoprolo, Methotrexate, and Rifampicin. UPDDI Dillon Gavlock 4 2019-07-16 Taylor_EPA 121
View/Edit EPA 4 2016-10-13 TOX SQL-SAL 1.5 Fluidic-3D -No Diseases- 356 0 0 0 0 0 0 Eighteen day study assessing toxicity of Erythromycin, Famotidine, Levofloxacin, Rosglitazone, and Trovafloxcin. UPDDI Dillon Gavlock 4 2019-07-16 Taylor_EPA 122
View/Edit Tier1_Lonza 2017-01-28 CC Kidney Proximal Tubule [384-well Static], Kidney Proximal Tubule [Fluidic] Fluidic-3D, Static-2D -No Diseases- 2,480 0 1,026 0 0 0 4 This study includes 38 fluidic MPS chips, and 168 static wells (384-well plate) seeded with Lonza renal proximal tubule (RPTEC) cells. The study includes responses to pH change (ammoniagenesis) and also Vitamin D metabolism. At endpoint, chips were either imaged for Live/Dead viability, or sent for genomic testing. Study characterizes: 1 - Effect of polymixin B on cell viability, KIM-1 expression, and transcriptome profile 2 - Ammoniagenesis response to changes in pH 3 - Metabolism of 25-OH vitamin D3 to (1a,25-(OH)2-vitD3) and (24,25-(OH)2-vitD3) in the presence of vehicle and calcitriol. TEX-VAL Luke Bergenthal 4 2018-06-27 Rusyn_TAMU 131
View/Edit Tier1_HIM31 2017-02-07 CC Kidney Proximal Tubule [384-well Static], Kidney Proximal Tubule [Fluidic] Fluidic-3D, Static-2D -No Diseases- 1,747 0 137 0 0 0 4 TAMU Kidney Proximal Tubule 10 chip study of fluidic devices with HIM-31 cells from the University of Washington. Study includes response to Polymyxin B, pH change and also Vitamin D metabolism. Some chips in the study were imaged for Live/Dead and others for sent for genomic testing. Study characterizes: 1 - Effect of polymixin B on cell viability, KIM-1 expression, and transcriptome profile 2 - Response to changes in pH through ammoniagenesis 3 - Metabolism of 25-OH vitamin D3 to (1a,25-(OH)2-vitD3) and (24,25-(OH)2-vitD3) in the presence of vehicle and calcitriol. These chip studies were carried out side by side with HIM-31 cells grown in a 384-well static culture format. TEX-VAL Michael Castiglione 4 2017-10-05 Rusyn_TAMU 132
View/Edit NVU_20170307 2017-03-07 CC Blood-Brain Barrier (NVU) Fluidic-3D -No Diseases- 1,289 0 0 0 0 0 1 March 7th, 2017 TAMU Blood-Brain Barrier (NVU) study. The purpose of this study was to demonstrate the creation of a functional blood-brain barrier (BBB) using the Vanderbilt University Neurovascular Unit (NVU). Barrier formation was determined using daily measurements of FITC-labeled 10kDa dextran. Each NVU device has media flowing through the device at a constant 2uL/min from two inlet ports supplied by syringe pump. The vascular side of the device was continuously perfused with EBM-2 media containing 1mg/mL FITC-dextran. The brain side compartment of the NVU was perfused with growth medium without FITC-dextran. Effluents were collected over a one hour period each day from both the vascular and brain sides of the device. The amount of FITC-dextran was determined by measuring the fluorescence in a 50uL aliquot of each effluent. As the endothelial cells proliferate and form tight junctions creating the BBB, one observed a decrease in the amount of FITC-dextran appearing in the effluent collected from the brain side of the NVU. All of the NVUs demonstrated some degree of FITC-dextran exclusion from the brain side effluent with increasing time in culture. Of the nine NVUs started for this study, four remained operational throughout the study period and exhibited barrier formation. After 24days in culture the barrier function on the four remaining NVUs was testing using Terfenadine and Fexofenadine. Terfenadine, a drug with know CNS effects, was expected to cross the NVU's BBB and be detectable in the brain side effluent. Fexofenadine, a drug without CNS effects and the active metabolite of Tefenadine, was expected to remain only in the vascular side effluent. Drugs were introduced from the vascular side in growth media that also contained 1mg/mL FITC-dextran. NVUs were exposed to drug for 2hr during which effluent was collected from both the vascular and brain side compartments. This was a dose escalation study where the first dose tested was 0.01uM. After 2hr, flow was stopped, new syringes were attached to the vascular side inlet of the NVU with the next higher concentration of drug. A total of 4 concentrations were tested (0.01, 0.1, 1 and 10uM) over a single 8hr period. A single NVU was included as a vehicle control. Both Terfenadine and Fexofenadine were prepared as 50mM stocks in DMSO. The drugs were serially diluted in growth media for testing and the vehicle control was tested using DMSO equivalent to the 10uM drug (a 1:5000 diultion). Effluent samples were tested for the presence of FITC-dextran and lactate dehydrogenase release as surrogate markers for compromised blood-brain barrier integrity and cellular viability, respectively. TEX-VAL Michael Castiglione 4 2017-11-06 Rusyn_TAMU 133
View/Edit Tier2.1_Lonza 2017-05-08 TOX Kidney Proximal Tubule [Fluidic] Fluidic-3D -No Diseases- 1,319 0 171 0 0 0 2 Kidney Promixal Tubule compound toxicity study. In this study, 24 fluidic chips were perfused with the test compound until significant disruption of the tubule was observed. The effects of compounds Cisplatin (6.4, and 64uM), Gentamicin (200, and 600uM), and Cadmium-II Chloride (0.05, and 0.5uM) on KIM-1 production and LDH release were studied. This study ended with endpoint/terminal assays of immunocytochemistry and transcriptomics. Note: This study is the fluidic half of the Tier 2 that had to be re-run due to issues with devices in the original run. TEX-VAL Michael Castiglione 4 2018-08-01 Rusyn_TAMU 135
View/Edit Tier2_Enteroid 2018-02-19 TOX Intestinal Enteroid [Monolayer TW], Intestinal Enteroid [TAMU Caco-2 TW] Fluidic-3D, Static-3D -No Diseases- 6,024 0 0 0 0 0 6 TAMU/IBT Tier-2 Human gut study. The overall study plan is a comparison of normal human jejunal-derived enteroids from the laboratory of Dr. Mary Estes at the Baylor College of Medicine with the colorectal adenocarcinoma derived Caco-2 cell line (HTB-37™) obtained from the ATCC that is considered an industry standard for drug permeability studies. The model system used for these studies is a two-dimensional transwell into which the enteroids and Caco2 cells are seeded. Over a period of 7 days in differentiation media the jejunal enteroids form a confluent monolayer with defined apical and basolateral surfaces, develop tight junctions and form a functional barrier. Likewise, over a period of 21 days differentiation after plating, the Caco-2 cells also form a confluent monolayer with defined apical and basolateral surfaces, develop tight junctions and form a functional barrier. Tier-2 testing focuses on extending the use of the microphysiological systems. The testing center greatly expanded the number of drugs being tested using both the enteroid and Caco-2 model system. A set of 12 drugs with well-characterized Caco-2 permeability (Optimization of the Caco-2 permeability assay to screen drug compounds for intestinal absorption and efflux. Press B. Methods Mol Biol. 2011;763:139-54. doi: 10.1007/978-1-61779-191-8_9. PMID:21874449) were used to establish a permeability standard curve. The drugs were then tested in the enteroid model to determine whether a similar curve could be constructed. These standard curves were then used to allow for the testing of an additional 13 drugs recommended for testing by the IQ Consortium. As with Tier-1 testing, the development and maintenance of barrier integrity were measured using transepithelial electrical resistance (TEER). The maintenance of barrier function during drug treatment was determined using both TEER (before and after drug exposure) and by the movement of 10kDa-FITC-labeled dextran from the comparment containing the drug to the receiving side. Two drug concentrations were tested, 10µM and 1µM for all drugs used in the Tier-2 studies. For permeability studies, the drugs were added on the apical side of the cell barrier. After 90 minutes at 37ºC in the cell culture incubator, aliquots of the media were removed from both the apical and basolateral compartments of the transwell device. The amount of 10kDa-FITC-labeled dextran in the media was quantified using a fluorescent plate reader. The viability of the cells in the transwells after drug treatment was measured using CellTiter Glo from Promega. The drug levels in the media samples were quantified using LC/MS. Additional studies were performed to measure drug efflux pathways by adding drugs to the basolateral side of the transwell and then quantifying the amount of drug in media samples taken from both the basolateral and apical sides of the device. TEX-VAL Michael Castiglione 4 2019-12-01 Rusyn_TAMU 138
View/Edit Tier1_3D_Bone 2017-08-08 TOX-DM-CC Bone Static-3D -No Diseases- 1,251 0 48 0 0 0 6 In this study, 96 scaffolds were shipped to TAMU from the developer lab. 48 of the bone scaffolds contained only human osteoblasts (TE-Bone), and 48 of the scaffolds contained both human osteoblasts as well as Ewing's sarcoma cell aggregates (TE-Tumor). These TE-Bone and TE-Tumor scaffolds were treated with 4.16uM Doxorubicin, 12uM Linsitinib, or their associated controls (0.04% or 0.12% DMSO, respectively). 3D scaffolds were treated in a realistic regimen over either 2 weeks (Doxorubicin) or 3 weeks (Linsitinib). Media was collected daily and tested for osteopontin (OPN) and LDH. At endpoint, scaffolds were either lysed for viability testing (total luminescence), or fixed and sectioned for immunocytochemistry (H&E). TEX-VAL Admin Sandra Karcher 4 2019-01-04 Rusyn_TAMU 139
View/Edit Heart_Tier1_CDI_2D 2018-01-01 TOX-CC Cardiac Static [TAMU 384w], Cardiac Static [TAMU 96w] Static-2D -No Diseases- 1,120 0 222 22 0 0 2 This tier of testing was a 2D companion to the 3D heart chip study to measure the effects of drugs on cardiomyocytes seeded within the UC-Berkeley chips. In this study, 176 wells were seeded with iPSC-derived cardiomyocytes purchased from CDI in a 384-well cell culture plate. Cells were cultured over 4 days in the plate prior to testing with reference compounds. Cultures were treated with either a media control (n=16), vehicle (0.1% DMSO, n=10), cisapride, verapamil, or isoproterenol for a 25 minute exposure at concentrations of 1, 10, 100, 1000, and 10000nM (n=10 for each condition). After each exposure, phase contrast images were captured (10FPS) from 3 wells in each condition, and analyzed using the MotionGUI script in Matlab (provided by UC-Berkeley). The outputs of this program were beat rate (BPM), time interval (s, duration between contraction and relaxation), contraction velocity (px/s), and relaxation velocity (px/s). The other wells were treated with a calcium sensitive dye, and measured for calcium flux over time with outputs of beats/min, and decay/rise ratio. Lastly, an additional 96-well plate was seeded for immunocytochemistry (DNA, actin filaments) at endpoint. TEX-VAL Admin Sandra Karcher 4 2019-09-30 Rusyn_TAMU 140
View/Edit Tier2_Lonza_Static 2017-03-01 TOX Kidney Proximal Tubule [384-well Static] Static-2D -No Diseases- 839 0 71 0 0 0 3 Companion study to the 3D, micro-fluidic study Tier 2 Lonza study. This study includes 72 static wells from a 384 well plate, seeded with Lonza renal proximal tubule cells (RPTECs). In this study, cells are treated with nephrotoxic compounds (cisplain, gentamicin, and cadmium) to study their effects on a monolayer. TEX-VAL Michael Castiglione 4 2018-06-27 Rusyn_TAMU 141
View/Edit KPT Tier0 Drug Binding 2017-06-15 CC Kidney Proximal Tubule [No Cells], RED - Kidney Proximal Tubule Fluidic-3D, Static-2D -No Diseases- 550 0 0 0 0 0 2 There are two parts in the Tier0 study for the proximal tubule model. The first part looked at non-specific chip binding. Blank (no cells) proximal tubule chips (Nortis Single Channel HARV1) were coated with ECM, then perfused with the treatment compounds used in the Tier2 Lonza study. Effluent concentrations are compared against stock solutions to determine non-specific chip binding. The second part determined the free faction of the compounds used in the Tier2 Lonza study through the use of rapid equilibrium dialysis (RED) plates. This fraction was calculated for the media used in the proximal tubule model—Lonza cell culture REGM (renal epithelial growth media). Non-Specific Chip Binding Cadmium: 9 chips (3 each at 0.05, 0.5, or 5µM ) Cisplatin: 10 chips (5 each at 6.4 or 64µM) Gentamicin was not run RED Free Fraction Cadmium: 12 samples (6 each at 0.05 or 0.5µM ) Cisplatin: 12 samples (6 each at 6.4 or 64µM) Gentamicin: 16 samples (8 each at 200 or 600µM) TEX-VAL Admin Sandra Karcher 4 2018-08-01 Rusyn_TAMU 142
View/Edit Tier2_3D_Bone 2017-10-27 TOX-DM Bone Static-3D -No Diseases- 4,738 0 66 0 0 0 4 In this study, 96 scaffolds were shipped to TAMU from Columbia University. All of the tissue scaffolds contained both human osteoblasts as well as Ewing's sarcoma cell aggregates (TE-Tumor). These TE-Tumor scaffolds were treated with control media, cisplatin (3, 10, and 30 uM), dexamethasone (0.1, 1, and 10 uM), methotrexate (0.1, 1, and 10 uM), vincristine (0.1, 1, and 10 uM), or MAP (combination of cisplatin, methotrexate and doxorubicin at a high and low exposure concentration). 3D scaffolds were treated in a realistic regimen over 16 days, then allowed 2 weeks to recover prior to endpoint analyses. Media was collected daily and tested for osteopontin (OPN), LDH, and the concentration of drug treatment (to determine recovery from stocks after treatment). At endpoint, scaffolds were either lysed for viability testing (total luminescence), or fixed and sectioned for immunocytochemistry (H&E). TEX-VAL Admin Sandra Karcher 4 2019-01-04 Rusyn_TAMU 143
View/Edit Tier1a_Upitt_3D 2018-04-19 CC SQL-SAL 1.5 Fluidic-3D -No Diseases- 581 0 36 0 0 0 1 This tier of testing was part of the 3D “Baseline” study to measure the secretome of cells seeded in the SQL-SAL tissue chip without any treatments. In this study, 6 chips (Nortis, Inc.) were seeded with primary hepatocytes from ThermoFisher, as well as supporting cells (LX-2, EA.Hy926, and THP-1) in a layered collagen matrix. Cells were cultured over 11 days with daily media sampling. Media was tested for flow rate, Albumin, Urea-Nitrogen, LDH, and TNF-a. TEX-VAL Admin Sandra Karcher 4 2019-07-11 Rusyn_TAMU 147
View/Edit Tier1a_Upitt_2D 2018-04-19 CC SQL-SAL [TAMU 96w] Static-2D -No Diseases- 528 0 205 0 0 0 2 This tier of testing was part of the 2D “Baseline” study to measure the secretome of cells plated in 2D culture without any treatments. In this study, 50 wells from a 2D cell culture plate (96-well plate) were plated in one of two configurations: with (n=30), or without (n=20) a collagen layer over the top of the cell monolayer. Wells were plated with primary hepatocytes from ThermoFisher, as well as supporting cells (LX-2, EA.Hy926, and THP-1). Cells were cultured over 10 days with daily media sampling. Media was tested for Albumin, Urea-Nitrogen, LDH, and TNF-a. It was determined that cells behaved better without the collagen layer on the top, so further 2D studies were carried out without the collagen layer over the cells. TEX-VAL Admin Sandra Karcher 4 2019-07-11 Rusyn_TAMU 148
View/Edit Tier1_2D_Bone 2017-09-08 TOX-DM-CC Bone [TAMU Static] Static-2D -No Diseases- 289 0 40 0 0 0 5 In this study, Ewing's sarcoma cells provided by the developer lab were cultured in a monolayer in 96 well plates. This study contains 2 plates with 80 wells on each (total of 160 wells across the study), treated with either Doxorubicin (4.16uM) or Linsitinib (12uM), and their relative controls. Media was sampled daily and tested for LDH release, and endpoints consisted of either tumor cell viability (luminescence assay), or phase contrast imaging. Lysates were also collected for transciptomics analysis. TEX-VAL Michael Castiglione 4 2018-06-28 Rusyn_TAMU 149
View/Edit Tier2_2D_Bone 2017-10-24 TOX-DM Bone [TAMU Static] Static-2D -No Diseases- 948 0 216 0 0 0 4 In this study, Ewing's sarcoma cells provided by the developer lab were cultured in a monolayer in 96 well plates. This study contains 4 plates with a total of 216 wells across the study, treated with control media, cisplatin (3, 10, and 30 uM), dexamethasone (0.1, 1, and 10 uM), methotrexate (0.1, 1, and 10 uM), vincristine (0.1, 1, and 10 uM), or MAP (combination of cisplatin, methotrexate and doxorubicin at a high and low exposure concentration). Media was sampled daily and tested for LDH release, and endpoints consisted of either tumor cell viability (luminescence assay), or phase contrast imaging. Lysates were also collected for transciptomics analysis. TEX-VAL Michael Castiglione 4 2019-01-04 Rusyn_TAMU 150
View/Edit TCTC Training 2018-02-27 TOX SQL-SAL 1.5 Fluidic-3D -No Diseases- 126 0 0 0 0 0 0 Purpose: The goal of this experiment was to educate Courtney Sakolish from Texas A&M University on the construction and use of the SQL-SAL 1.5 liver model in a commercial Nortis devices. Methods: The SQL-SAL 1.5 liver model was constructed in 6 Nortis devices for 14 day treatment to vehicle control or 88 and 220 μM tolcapone. Results: Albumin, urea synthesis was decreased, LDH release was increased in devices treated to tolcapone as compared to vehicle only treated devices. UPDDI Dillon Gavlock 4 2018-09-18 Taylor_MPS 152
View/Edit Tier1_Enteroid 2017-09-07 CC Intestinal Enteroid [Monolayer TW], Intestinal Enteroid [TAMU Caco-2 96w], Intestinal Enteroid [TAMU Caco-2 TW], Intestinal Enteroid [TAMU J2 96w] Fluidic-3D, Static-2D, Static-3D -No Diseases- 5,120 0 0 0 0 0 3 TAMU/IBT Tier-1 Human gut study. The overall study plan is a comparison of normal human jejunal-derived enteroids from the laboratory of Dr. Mary Estes at the Baylor College of Medicine with the colorectal adenocarcinoma derived Caco-2 cell line (HTB-37™) obtained from the ATCC that is considered an industry standard for drug permeability studies. The model system used for these studies is a two-dimensional transwell into which the enteroids and Caco2 cells are seeded. Over a period of 7 days in differentiation media, the jejunal enteroids form a confluent monolayer with defined apical and basolateral surfaces, develop tight junctions and form a functional barrier. Likewise, over a period of 21 days differentiation after plating, the Caco-2 cells also form a confluent monolayer with defined apical and basolateral surfaces, develop tight junctions and form a functional barrier. Tier-1 testing focuses on testing drugs that the tissue chip developers have already tested in their models as a way to evaluate 1) the successful transfer of the model system from the developer's laboratory to the tissue chip testing center's laboratory and 2) examine the reproducibility of results. In addition to testing the transfer and reproducibility of the human jejunal enteroid model, the tissue chip testing center is using the Caco-2 model as a comparator. For these studies, the development and maintenance of barrier integrity were measured using transepithelial electrical resistance (TEER). The maintenance of barrier function during drug treatment was determined using both TEER (before and after drug exposure) and by the movement of 10kDa-FITC-labeled dextran from the compartment containing the drug to the receiving side. Two drug concentrations were tested, 10µM and 1µM for all drugs used in the Tier-1 studies. For permeability studies, the drugs were added on the apical side of the cell barrier. After 90 minutes at 37ºC in the cell culture incubator, aliquots of the media were removed from both the apical and basolateral compartments of the transwell device. The amount of 10kDa-FITC-labeled dextran in the media was quantified using a fluorescent plate reader. The viability of the cells in the transwells after drug treatment was measured using CellTiter Glo from Promega. The drug levels in the media samples were quantified using LC/MS. Additional studies were performed to measure drug efflux pathways by adding drugs to the basolateral side of the transwell and then quantifying the amount of drug in media samples taken from both the basolateral and apical sides of the device. Also during Tier-1 testing, the testing center examined the concentration dependence of drug-induced cytotoxicity in the Caco-2 cells grown as a two-dimensional monolayer of cells in a standard tissue culture plate. Similar cytotoxicity studies were performed using the enteroids plated on collagen-coated wells. Because of the short time to differentiate the enteroids, 5 days compared with 21 days for Caco2 cells, drug cytotoxicity was tested in both undifferentiated enteroids and enteroids grown for 5-7 days in differentiation media. TEX-VAL Michael Castiglione 4 2019-12-01 Rusyn_TAMU 156
View/Edit 1700500003 2017-10-24 TOX Brain Spheroids Fluidic-3D -No Diseases- 36 0 0 0 0 0 1 Dose escalation study of Docetaxel in the Brain Spheroid/Mini-Brain MPS. The measured outputs were LDH, DJ-1, and NAA (1-Naphthaleneacetic acid) TC2T Michael Castiglione 4 2019-07-17 Cirit_MIT 157
View/Edit 1700500004 2017-10-24 CC Brain Spheroids Fluidic-3D -No Diseases- 117 0 0 0 0 0 1 Dose response study of Bortezomib, Tamoxifen, Doxycycline, and Vincristine. Measured outputs are LDH, DJ-1, and NAA (1-Naphthaleneacetic acid) TC2T Michael Castiglione 4 2019-07-16 Cirit_MIT 158
View/Edit Tier1b_Upitt_3D 2018-05-23 CC SQL-SAL 1.5 Fluidic-3D -No Diseases- 858 0 36 0 0 0 2 This tier of testing was part of the 3D “Metabolism” study to measure the metabolic capacity of cells seeded in SQL-SAL tissue chips. In this study, 8 chips (Nortis, Inc.) were seeded with primary hepatocytes from ThermoFisher, as well as supporting cells (LX-2, EA.Hy926, and THP-1) in a layered collagen matrix. Cultures were treated with either a vehicle (0.1% DMSO, n=4), or a 10uM Terfenadine (n=4). Cells were cultured over 13 days with daily media sampling. Media was tested for flow rate, Albumin, Urea-Nitrogen, LDH, and TNF-a, as well as the presence of the parent compound (terfenadine), and the major metabolite (fexofenadine). Additionally, viability testing was carried out at endpoint. Note that there was a power outage that stalled syringe pumps on days 11-12. TEX-VAL Michael Castiglione 4 2019-07-26 Rusyn_TAMU 161
View/Edit Tier1b_Upitt_2D 2018-05-23 CC SQL-SAL [TAMU 96w] Static-2D -No Diseases- 750 0 60 0 0 0 1 2D Companion experiment to Tier1b_Upitt_3D This tier of testing was part of the 2D “Metabolism” study to measure the metabolic capacity of cells plated in 2D culture. In this study, 20 wells from a 2D cell culture plate (96-well plate) were plated with primary hepatocytes from ThermoFisher, as well as supporting cells (LX-2, EA.Hy926, and THP-1). Cultures were treated with either a vehicle (0.1% DMSO, n=10), or a 10uM Terfenadine (n=10). Cells were cultured over 13 days with daily media sampling. Media was tested for Albumin, Urea-Nitrogen, LDH, and TNF-a, as well as the presence of the parent compound (terfenadine), and the major metabolite (fexofenadine). Additionally, viability testing was carried out at endpoint TEX-VAL Michael Castiglione 4 2019-07-11 Rusyn_TAMU 162
View/Edit 1700400006 2017-10-30 CC Intestinal Enteroid [Monolayer TW] Fluidic-3D -No Diseases- 82 0 0 0 0 0 1 MIT Tier-1 Enteroid MPS Study This study tested the effect of a one day dose of 10 µM diclofenac, fexofenadine, or terfenadine on JHU's Enteroid transwell monolayer MPS model. Transwell plates were seeded with 50 J212 enteroid fragments, and allowed to form a monolayer. The development and maintenance of barrier integrity was measured using transepithelial electrical resistance (TEER) both before and after the one day of compound addition. Cell function was assayed by measuring the amount of secreted fatty acid-binding protein 2 (FABP2). TC2T Michael Castiglione 4 2018-07-17 Cirit_MIT 164
View/Edit Estrogen Dependent Growth of MCF7 Cells 2017-01-11 DM LAMPS MCF7 Metastatic Breast Cancer Model Fluidic-3D -No Diseases- 42 0 90 0 0 0 0 LAMPS model to examine growth of MCF7 ESR1 mutants (WT, Y537S, D538G) in the presence or absence of Estrogen. 6 total chips, over 17 days, quantifying change in fluorescent expression of the mutant cell types over time. While both mutants (Y537S and D538G) demonstrate increased constitutive growth compared to WT in the absence of estrogen, the Y537S muatnt shows enhanced estrogen dependent growth compared to the D538G mutant. This is similar to observations made in co-culture models, but different to phenotypes observed in monoculture where the D538G mutant shows estrogen-enhanced growth. UPDDI Dillon Gavlock 4 2020-02-17 Taylor_MPS 165
View/Edit Estrogen Dependent Growth of MCF7 Cells 2 2018-02-18 DM LAMPS MCF7 Metastatic Breast Cancer Model Fluidic-3D -No Diseases- 276 0 180 0 0 0 0 LAMPS model to examine growth of MCF7 ESR1 mutants. 12 total chips, over 17 days, quantifying change in fluorescent expression of the mutant cell types over time. Oxygen tension was varied from 15 ul/hr and 5 ul/hr. The goal was to examine the effects of changes in oxygen tension on growth phenotypes conferred by ESR1 mutant-expressing cells in the LAMPS models. Whereas monoculture studies performed at zone 3 (3-6%) mimicked the results obtained at zone 1, both co-culture and LAMPS models performed at low oxygen showed distinct differences compared to studies performed at zone 1 oxygen tension. While the constitutive growth advantage for ESR1 mutants remained unaffected, the enhanced E2-dependent growth for the Y537S mutation and the E2-dependent growth for WT expressing cells were lost. Overall, these results demonstrate the importance of oxygen tension in regulating specific growth phenotypes conferred by ESR1 mutants. UPDDI Dillon Gavlock 4 2020-02-17 Taylor_MPS 166
View/Edit Estrogen Dependent Growth of MCF7 Cells 3 2018-03-21 DM LAMPS MCF7 Metastatic Breast Cancer Model Fluidic-3D -No Diseases- 138 0 90 0 0 0 0 LAMPS model to examine growth of MCF7 ESR1 mutants. 12 total chips, over 17 days, quantifying change in fluorescent expression of the mutant cell types over time. Oxygen tension: 5 ul/hr. The goal was to examine the effects of changes in oxygen tension on growth phenotypes conferred by ESR1 mutant-expressing cells in the LAMPS models. Whereas monoculture studies performed at zone 3 (3-6%) mimicked the results obtained at zone 1, both co-culture and LAMPS models performed at low oxygen showed distinct differences compared to studies performed at zone 1 oxygen tension. While the constitutive growth advantage for ESR1 mutants remained unaffected, the enhanced E2-dependent growth for the Y537S mutation and the E2-dependent growth for WT expressing cells were lost. Overall, these results demonstrate the importance of oxygen tension in regulating specific growth phenotypes conferred by ESR1 mutants. UPDDI Dillon Gavlock 4 2019-08-13 Taylor_MPS 167
View/Edit Effect of AZD9496 on growth of MCF7 Y537S Mutant Cells 2018-07-10 DM LAMPS MCF7 Metastatic Breast Cancer Model Fluidic-3D -No Diseases- 56 0 0 0 0 0 0 The goal was to examine the effect of the ER antagonist AZD9496 on growth of MCF7 Y537S Mutant Cells in LAMPS. While the Y537S mutant shows resistance to AZD9496 treatment in monoculture, this mutant does not show significant resistance to treatment with this drug in the LAMPS model, suggesting the Y537S mutant is more sensitive to this ER antagonist in the liver microenvironment. UPDDI Dillon Gavlock 4 2019-08-13 Taylor_MPS 171
View/Edit Vascularized Tumor-Tier1-2D 2018-06-12 TOX-EFF-CC Vascularized Tumor [TAMU 96w] Static-2D -No Diseases- 260 0 0 0 0 0 4 Viability of endothelial cells and cancer cells mono-culture in a 2D 96-well plate after exposure to the drugs tested in the 3D platform. TEX-VAL Michael Castiglione 4 2019-11-13 Rusyn_TAMU 174
View/Edit 1800900004 2018-05-17 TOX-CC Skin MPS, Skin MPS [TW-12] Fluidic-3D, Static-3D -No Diseases- 63 0 0 0 0 0 1 MIT study of the Columbia Skin MPS. Constructs created at Columbia and shipped ready-to-use to MIT TCTC. This study contains constructs in the HeLiVaSkCa Skin Module device and in a 12-well transwell plate. This experiment tested the effect of a two day dose of doxorubicin (10, 1, or 0.1 µM) by measuring LDH production and PrestoBlue metabolism in 12 HeLiVaSkCa constructs. The four transwell constructs were untreated. TC2T Michael Castiglione 4 2018-07-17 Cirit_MIT 179
View/Edit Tier0 Bone 2017-09-04 CC Bone, RED - Bone Static-2D, Static-3D -No Diseases- 1,938 0 0 0 0 0 2 In Tier 0, 42 blank wells (HeLiVaSkCa Bone Cancer Module) were treated with the compounds and concentrations used in Tiers 1 and 2 of the TE-Bone/Tumor study. The purpose of this study was to determine platform binding of these treatment compounds. Blank wells were treated with cisplatin (3, 10, 30uM), dexamethasone (0.1, 1, 10uM), linsitinib (12um), methotrexate (0.1, 1, 10uM), or vincristine (0.1, 1, 10uM) over 21 days, or doxorubicin (4.16uM) over 14 days following the specific on/off treatment regimen for each compound. Media samples were collected daily and compared against stock solutions to determine % recovery from the platforms. Additionally, rapid equilibrium dialysis (RED) plates were used to determine the free fraction of these compounds in the cell culture media at each of these concentrations. TEX-VAL Michael Castiglione 4 2019-02-03 Rusyn_TAMU 182
View/Edit TCTC training MIT 2018-05-02 TOX SQL-SAL 1.5 Fluidic-3D -No Diseases- 164 0 0 0 0 0 1 Purpose: The goal of this experiment was to educate Michael Shockley from MIT on the construction and use of the SQL-SAL 1.5 liver model in a commercial Nortis devices. Methods: The SQL-SAL 1.5 liver model was constructed in 6 Nortis devices for 14 day treatment to vehicle control or 88 and 220 μM tolcapone. Results: Albumin, urea synthesis was decreased, LDH release was increased in devices treated to tolcapone as compared to vehicle only treated devices. UPDDI Richard DeBiasio 4 2019-07-16 Taylor_MPS 184
View/Edit 1800700001 2018-01-31 CC Bone Static-3D -No Diseases- 216 0 0 0 0 0 1 Initial experiment with Columbia's Bone-Tumor model. Twelve constructs were created as CU and then shipped to MIT in one HeLiVaSkCa Bone Cancer Module plate. Three of those constructs were treated with 250µM Vincristine for 4 days, while the remaining 8 constructs were untreated. Constructs' LDH and bone bio-marker production were measured over seven days. After seven days, end point measurements of luciferase expression and alkaline phosphatase production were taken. TC2T Michael Castiglione 4 2019-07-10 Cirit_MIT 185
View/Edit 1800700006 2018-03-05 TOX Bone Static-3D -No Diseases- 477 0 0 0 0 0 1 Twelve constructs were created at CU and then shipped to MIT in one HeLiVaSkCa Bone Cancer Module plate. Three constructs each were treated with low, medium, or high dose (5/40/3nM; 50/300/30nM; 500/4000/300nM) of vincristine, cyclophosphamide, and doxorubicin (VCD) , while the remaining three constructs were untreated. Constructs' LDH, alkaline phosphatase, and bone biomarker production were measured over seven days, with treatment starting at Day 0. After six days, the cells within the constructs were lysed, and an end point measurement of luciferase expression was taken. Biomarker profiles of sclerostin, osteoprotegrin, and osteonectin are most likely related to tumor viability, as decreased secretion was related to a higher drug-dose. Secreted osteopontin did not follow a dose-dependent or tumor viability-related trend. TC2T Michael Castiglione 4 2019-07-10 Cirit_MIT 187
View/Edit Tier1c_Upitt_3D 2018-06-23 TOX-EFF SQL-SAL 1.5 Fluidic-3D -No Diseases- 2,696 0 72 0 0 0 2 This tier of testing was part of the 3D “Toxicity” study to measure the effects of drugs on cells seeded in SQL-SAL tissue chips. In this study, 36 chips (Nortis, Inc.) were seeded with primary hepatocytes from ThermoFisher, as well as supporting cells (LX-2, EA.Hy926, and THP-1) in a layered collagen matrix. Cultures were treated with either a vehicle (0.1% DMSO, n=6), caffeine (600uM, n=4), trovafloxacin (150uM, n=5), troglitazone (28uM, n=6), tolcapone (88uM, n=3), rosiglitazone (0.8uM, n=3), pioglitazone (3uM, n=3), trovaflocacin + LPS (150uM + 1ug/mL, n=3), or LPS (1ug/mL, n=3). Cells were cultured over 10 days with daily media sampling. Media was tested for flow rate, Albumin, Urea-Nitrogen, LDH, and TNF-a, as well as the presence of the parent drug compounds. Additionally, viability testing was carried out at endpoint. Note: This study was carried out in 3 “arms” of testing: A, B, and C as noted on PTMs. TEX-VAL Michael Castiglione 4 2019-07-11 Rusyn_TAMU 189
View/Edit Heart_Tier1_UCB_3D 2018-02-05 CC Cardiac MPS Fluidic-3D -No Diseases- 624 0 62 15 0 0 2 This tier of testing was part of the 3D heart chip study to measure the effects of drugs on cardiomyocytes seeded within the UC-Berkeley chips. In this study, 10 chips were seeded with iPSC-derived cardiomyocytes provided by UC-Berkeley, as well as supporting fibroblasts (iPS-stromal cells) in a central growth chamber. Cells were cultured over 4 days in the chip prior to testing with reference compounds. Cultures were treated with either a vehicle (0.1% DMSO, n=3), cisapride (n=3), verapamil (n=3), or isoproterenol (n=4) in a dose escalation format with 25 minute exposures at concentrations of 1, 10, 100, 1000, and 10000nM. After each exposure, phase contrast images were captured (10FPS), and analyzed using the MotionGUI script in Matlab (provided by UC-Berkeley). The outputs of this program were beat rate (BPM), time interval (s, duration between contraction and relaxation), contraction velocity (px/s), and relaxation velocity (px/s). Immunohistochemistry was difficult in this chip, so images are displayed as phase contrast only. TEX-VAL Michael Castiglione 4 2019-09-27 Rusyn_TAMU 190
View/Edit 1801100001 2018-07-14 CC SQL-SAL 1.5 Fluidic-3D -No Diseases- 171 0 0 0 0 0 1 The first MIT Pitt Liver MPS experiment (1801100001 - healthy controls) TC2T Michael Castiglione 4 2019-07-10 Cirit_MIT 191
View/Edit MCF7 Co-Culture Plate Study 1 2017-08-09 DM LAMPS MCF7 Metastatic Breast Cancer Plate Model Static-3D -No Diseases- 114 0 72 0 0 0 0 The purpose of this study was to create a point of comparison between static and fluidic modeling of the LAMPS + MCF7 mutant model. While both mutants (Y537S and D538G) demonstrate increased constitutive growth compared to WT in the absence of estrogen, the Y537S muatnt shows enhanced estrogen dependent growth compared to the D538G mutant. This is similar to observations made in LAMPS models, but different to phenotypes observed in monoculture where the D538G mutant shows estrogen-enhanced growth. UPDDI Dillon Gavlock 4 2019-08-13 Taylor_MPS 195
View/Edit 1800700004 2018-03-20 CC Bone Tumor Spheroids [MIT] Fluidic-3D -No Diseases- 15 0 0 0 0 0 1 Tumor cell only characterization for the Bone-Tumor MPS model. Cells treated with 24 hours of V/C/D combination at low, medium, and high doses. TC2T Michael Castiglione 4 2019-07-11 Cirit_MIT 199
View/Edit 1800700005 2018-03-29 CC Bone Tumor Spheroids [MIT] Fluidic-3D -No Diseases- 27 0 0 0 0 0 1 Measure the luciferase response of untreated Bone-Tumor/Ewing's Sarcoma spheroids at different cell densities per tube. TC2T Michael Castiglione 4 2019-07-16 Cirit_MIT 200
View/Edit 1800300001 2018-01-16 TOX Brain, Brain MPS [MIT 96w Static] Static-2D, Static-3D -No Diseases- 1,478 0 0 0 0 0 2 Tier 2 UWM-NPC redo TC2T Michael Castiglione 4 2019-07-17 Cirit_MIT 201
View/Edit 1700300004 2017-05-16 TOX Brain MPS [MIT 96w Static] Static-2D -No Diseases- 736 0 0 0 0 0 1 2D Drug Screening with CDI Neurons TC2T Michael Castiglione 4 2019-07-16 Cirit_MIT 202
View/Edit 1700300002 2017-04-03 CC Brain MPS [MIT 96w Static] Static-2D -No Diseases- 297 0 0 0 0 0 1 2D Drug Screening with CDI neurons TC2T Michael Castiglione 4 2019-07-16 Cirit_MIT 203
View/Edit 1700300001 2017-03-01 CC Brain Static-3D -No Diseases- 70 0 0 0 0 0 1 UWM-NPC Homemade ECM vs StemPharm ECM TC2T Michael Castiglione 4 2019-07-16 Cirit_MIT 204
View/Edit Nortis Terfenadine Metabolism 2016-01-26 TOX-EFF SQL-SAL 1.5 Fluidic-3D -No Diseases- 150 0 0 0 0 0 0 Purpose: the goal of this experiment was to perfuse terfenadine at 6 and 3 µM into SQL-SAL 1.5 liver models to collect metabolism data for PBPK model development. Methods: Nortis devices were perfused with HMM media containing 6, 3 or 0 µM terfenadine. Media was collected at 8, 24 and 40 hours. Samples were extracted with acetonitrile for mass spec determination of terfenadine and the metabolite fexofenadine. Results: Steady state concentration of terfenadine was used to calculate apparent intrinsic clearance for in vitro in vivo extrapolation. UPDDI Larry Vernetti 4 2019-07-16 Taylor_MPS 206
View/Edit Heart_Tier1_CDI_3D 2018-05-18 CC Cardiac MPS Fluidic-3D -No Diseases- 768 0 96 12 0 0 2 This tier of testing was part of the 3D heart chip study to measure the effects of drugs on cardiomyocytes seeded within the UC-Berkeley chips. In this study, 16 chips were seeded with iPSC-derived cardiomyocytes purchased from CDI (iCell Cardiomyocytes, lot #1293677) in a central growth chamber. Cells were cultured over 4 days in the chip prior to testing with reference compounds. Cultures were treated with either cisapride (n=3), verapamil (n=6), or isoproterenol (n=7) in a dose escalation format with 25 minute exposures at concentrations of 1, 10, 100, 1000, and 10000nM. After each exposure, phase contrast images were captured (10FPS), and analyzed using the MotionGUI script in Matlab (provided by UC-Berkeley). The outputs of this program were beat rate (BPM), time interval (s, duration between contraction and relaxation), contraction velocity (px/s), and relaxation velocity (px/s). Immunohistochemistry was difficult in this chip, so images are displayed as phase contrast only. TEX-VAL Michael Castiglione 4 2019-09-27 Rusyn_TAMU 208
View/Edit Heart_Tier1_UCB_2D 2018-02-05 CC Cardiac Static [TAMU 384w] Static-2D -No Diseases- 516 0 86 22 0 0 2 This tier of testing was a 2D companion to the 3D heart chip study to measure the effects of drugs on cardiomyocytes seeded within the UC-Berkeley chips. In this study, 43 wells were seeded with iPSC-derived cardiomyocytes provided by UC-Berkeley, as well as supporting fibroblasts (iPS-stromal cells) in a 384-well cell culture plate. Cells were cultured over 4 days in the plate prior to testing with reference compounds. Cultures were treated with either a media control (n=7), vehicle (0.1% DMSO, n=6), cisapride, verapamil, or isoproterenol for a 25 minute exposure at concentrations of 1, 10, 100, 1000, and 10000nM (n=2 for each condition). After each exposure, phase contrast images were captured (10FPS), and analyzed using the MotionGUI script in Matlab (provided by UC-Berkeley). The outputs of this program were beat rate (BPM), time interval (s, duration between contraction and relaxation), contraction velocity (px/s), and relaxation velocity (px/s). TEX-VAL Michael Castiglione 4 2019-09-27 Rusyn_TAMU 209
View/Edit 1801100003 2018-08-03 TOX-CC SQL-SAL 1.5 Fluidic-3D -No Diseases- 240 0 0 0 0 0 1 MIT SQL-SAL 1.5 troglitazone study TC2T Michael Castiglione 4 2019-07-10 Cirit_MIT 211
View/Edit 1700100001 2017-02-03 CC Kidney Proximal Tubule [Fluidic] Fluidic-3D -No Diseases- 144 0 0 0 0 0 1 UWash Kidney Proximal Tubule model (Single Channel Nortis device version) KIM-1 and LDH production +/- two days of treatment with 50 µM Polymyxin B. Cell line used was UWash RPTEC donor cells (HIM-31). TC2T Michael Castiglione 4 2019-07-10 Cirit_MIT 212
View/Edit 1700100002 2017-02-09 CC Kidney Proximal Tubule [Fluidic] Fluidic-3D -No Diseases- 112 0 0 0 0 0 1 Attempted study of ammoniagenesis in UWash Kidney Proximal Tubule model with UWash RPTEC cells (HIM-31). Two chips were treated with 50 µM PMB for two days starting on day 12, while two chips were used as controls. KIM and LDH reading were taken every two days. Ammoniagenesis was measured on day 14. Unfortunately, the standard curve for the ammonia concentrations had an issue, so the ammoniagenesis of the model could not be quantified. TC2T Michael Castiglione 4 2019-07-10 Cirit_MIT 213
View/Edit 1700100003 2017-02-21 CC Kidney Proximal Tubule [Fluidic] Fluidic-3D -No Diseases- 216 0 0 0 0 0 1 This study measured effect of pH on ammoniagenesis in the Kidney Proximal Tubule model with either HIM-31 cells or Lonza cells. On day 10, six chips were treated with 6.9 pH buffer for eight hours, while five chips were treated with 7.4 pH buffer for the same time frame. Samples were taken every two hours, and their ammonia concentration were measured. On day 13, six chips were treated with 50µM polymyxin B for one day. KIM-1 samples were taken for all chips every two days for 14 days. TC2T Michael Castiglione 4 2019-07-10 Cirit_MIT 214
View/Edit 1700100006 2017-02-02 CC Kidney Proximal Tubule [No Cells] Fluidic-3D -No Diseases- 8 0 0 0 0 0 1 Cell free flow rate test for UWash Kidney Proximal Tubule model in four devices over two days. There was a dead-leg volume of 115µL. TC2T Michael Castiglione 4 2019-07-10 Cirit_MIT 215
View/Edit Hepatocyte Lot Testing 2018-10-11 CC SQL-SAL 1.5 Fluidic-3D -No Diseases- 250 0 0 0 0 0 1 The purpose of this lot testing is to define an adequate lot for usage in vLAMPS experiments moving forward. There will be 12 Nortis devices seeded with variable lots of hepatocytes (1838 (previous lot; used as control), 8135, 1951, 1960), 3 devices per lot. Three plates will be seed according to the plate figures in the supporting data and the Protocol file. The LAMPS 1.5 protocol will be used, the experiment will last 21 days, the following readout will be performed from sample efflux collected daily: Albumin, BUN, TNF-a, LDH. The following readouts will be collected from plate data: Lentiviral Transduction Efficiency, P450 activity in response to Rifampycin, Bile Canalicular Activity (CMFDA staining) UPDDI Dillon Gavlock 4 2019-08-13 Taylor_MPS 216
View/Edit Heart_Tier2_UCB_3D 2018-05-25 TOX Cardiac MPS Fluidic-3D -No Diseases- 4,104 0 511 58 0 0 2 This tier of testing was part of the 3D heart chip study to measure the effects of drugs on cardiomyocytes seeded within the UC-Berkeley chips. In this study, 55 chips were seeded with iPSC-derived cardiomyocytes provided by UC-Berkeley as well as supporting fibroblasts (iPS-stromal cells) in a central growth chamber. Cells were cultured over 5 days in the chip prior to testing with reference compounds. Cultures were treated with either vehicle (n=12, 0.1% DMSO), atenolol (0.01µM, n=4; 0.1µM, n=4; 1µM, n=3; 10µM, n=3), sorafenib (0.1µM, n=4; 1µM, n=4; 10µM, n=4; 100µM, n=3) or carbamazepine (0.1, 1, 10, 100µM, n=4 each) over a 1 week period. At each timepoint (baseline, 5 hours, and daily for days 1-7), phase contrast images were captured (10FPS), and analyzed using the MotionGUI script in Matlab (provided by UC-Berkeley). The outputs of this program were beat rate (BPM), time interval (s, duration between contraction and relaxation), contraction velocity (px/s), and relaxation velocity (px/s). Immunohistochemistry was difficult in this chip, so images are displayed as phase contrast only. TEX-VAL Michael Castiglione 4 2019-09-27 Rusyn_TAMU 217
View/Edit Heart_Tier2_UCB_2D 2018-09-06 TOX Cardiac Static [TAMU 384w] Static-2D -No Diseases- 3,640 0 433 0 0 0 2 This tier of testing was a 2D companion to the 3D heart chip study to measure the effects of drugs on cardiomyocytes seeded within the UC-Berkeley chips. In this study, 308 wells were seeded with iPSC-derived cardiomyocytes provided by UC-Berkeley, as well as supporting fibroblasts (iPS-stromal cells) in a 384-well cell culture plate. Cells were cultured over 5 days in the plate prior to testing with reference compounds. Cultures were treated with either a media control, vehicle (0.1% DMSO, atenolol (0.01, 0.1, 1, 10uM), sorafenib (0.1, 1, 10, 100uM), or carbamazepine (0.1, 1, 10, 100uM) over a week-long exposure (n=22 for each condition), measuring daily. At each timepoint, phase contrast images were captured (10FPS), and analyzed using the MotionGUI script in Matlab (provided by UC-Berkeley). The outputs of this program were beat rate (BPM), time interval (s, duration between contraction and relaxation), contraction velocity (px/s), and relaxation velocity (px/s). As an additional endpoint, cells were treated with a calcium sensitive dye on day 12, and calcium flux was measured as beats/min. TEX-VAL Michael Castiglione 4 2019-09-30 Rusyn_TAMU 218
View/Edit Tier1c_Upitt_2D 2018-08-04 TOX-CC SQL-SAL [TAMU 96w] Static-2D -No Diseases- 2,125 0 120 0 0 0 2 2D, 96-well based companion study for Tier1c_Upitt_3D This tier of testing was part of the 2D “Toxicity” study to measure the effects of drugs on cells plated in 2D culture. In this study, 60 wells from a 2D cell culture plate (96-well plate) were plated with primary hepatocytes from ThermoFisher, as well as supporting cells (LX-2, EA.Hy926, and THP-1). Cultures were treated with either a vehicle (0.1% DMSO, n=12), caffeine (600uM, n=6), trovafloxacin (150uM, n=6), troglitazone (28uM, n=6), tolcapone (88uM, n=6), rosiglitazone (0.8uM, n=6), pioglitazone (3uM, n=6), trovaflocacin + LPS (150uM + 1ug/mL, n=6), or LPS (1ug/mL, n=6). Cells were cultured over 10 days with daily media sampling. Media was tested for Albumin, Urea-Nitrogen, LDH, and TNF-a, as well as the presence of the parent drug compounds. Additionally, viability testing was carried out at endpoint. TEX-VAL Michael Castiglione 4 2019-07-11 Rusyn_TAMU 219
View/Edit UPLiver_Tier2a2b_3D 2018-08-15 CC SQL-SAL 1.5 Fluidic-3D -No Diseases- 1,248 0 26 0 0 0 2 This tier of testing was part of the 3D “Baseline” and “Metabolism” studies to measure the secretome and metabolic capacity of cells seeded in SQL-SAL tissue chips. In this study, 13 chips (Nortis, Inc.) were seeded with iPSC-derived hepatocytes from CDI, as well as supporting cells (LX-2, EA.Hy926, and THP-1) in a layered collagen matrix. Cultures were treated with either cell culture media (n=5), vehicle (0.1% DMSO, n=4), or a 10uM Terfenadine (n=4). Cells were cultured over 12 days with daily media sampling. Media was tested for Albumin, Urea-Nitrogen, LDH, and TNF-a, as well as the presence of the parent compound (terfenadine), and the major metabolite (fexofenadine). Additionally, viability testing was carried out at endpoint. Note that there was a power outage that stalled syringe pumps on days 11-12. TEX-VAL Michael Castiglione 4 2019-07-11 Rusyn_TAMU 223
View/Edit UPLiver_Tier2a2b_2D 2018-08-15 CC SQL-SAL [TAMU 96w] Static-2D -No Diseases- 888 0 60 0 0 0 2 2D companion to the UPLiver_Tier2a2b_3D Study This tier of testing was part of the 2D “Baseline” and “Metabolism” studies to measure the secretome and metabolic capacity of cells plated in 2D culture. In this study, 30 wells from a 2D cell culture plate (96-well plate) were plated with iPSC-derived hepatocytes from CDI, as well as supporting cells (LX-2, EA.Hy926, and THP-1). Cultures were treated with either cell culture media (n=10), vehicle (0.1% DMSO, n=10), or a 10uM Terfenadine (n=10). Cells were cultured over 12 days with daily media sampling. Media was tested for Albumin, Urea-Nitrogen, LDH, and TNF-a, as well as the presence of the parent compound (terfenadine), and the major metabolite (fexofenadine). Additionally, viability testing was carried out at endpoint. TEX-VAL Michael Castiglione 4 2019-07-11 Rusyn_TAMU 224
View/Edit UPLiver_Tier2c_3D 2018-08-31 TOX SQL-SAL 1.5 Fluidic-3D -No Diseases- 2,150 0 54 0 0 0 2 This tier of testing was part of the 3D “Toxicity” study to measure the effects of drugs on cells seeded in SQL-SAL tissue chips. In this study, 27 chips (Nortis, Inc.) were seeded with iPSC-derived hepatocytes from CDI, as well as supporting cells (LX-2, EA.Hy926, and THP-1) in a layered collagen matrix. Cultures were treated with either a vehicle (0.1% DMSO, n=3), caffeine (600uM, n=3), trovafloxacin (150uM, n=3), troglitazone (28uM, n=3), tolcapone (88uM, n=3), rosiglitazone (0.8uM, n=3), pioglitazone (3uM, n=3), trovaflocacin + LPS (150uM + 1ug/mL, n=3), or LPS (1ug/mL, n=3). Cells were cultured over 10 days with daily media sampling. Media was tested for flow rate, Albumin, Urea-Nitrogen, LDH, and TNF-a, as well as the presence of the parent drug compounds. Additionally, viability testing was carried out at endpoint. TEX-VAL Michael Castiglione 4 2019-07-11 Rusyn_TAMU 225
View/Edit UPLiver_Tier2c_2D 2018-08-31 TOX SQL-SAL [TAMU 96w] Static-2D -No Diseases- 1,946 0 120 0 0 0 2 2D companion to the UPLiver_Tier2c_3D Study This tier of testing was part of the 2D “Toxicity” study to measure the effects of drugs on cells plated in 2D culture. In this study, 60 wells from a 2D cell culture plate (96-well plate) were plated with iPSC-derived hepatocytes from CDI, as well as supporting cells (LX-2, EA.Hy926, and THP-1). Cultures were treated with either a vehicle (0.1% DMSO, n=12), caffeine (600uM, n=6), trovafloxacin (150uM, n=6), troglitazone (28uM, n=6), tolcapone (88uM, n=6), rosiglitazone (0.8uM, n=6), pioglitazone (3uM, n=6), trovaflocacin + LPS (150uM + 1ug/mL, n=6), or LPS (1ug/mL, n=6). Cells were cultured over 10 days with daily media sampling. Media was tested for Albumin, Urea-Nitrogen, LDH, and TNF-a, as well as the presence of the parent drug compounds. Additionally, viability testing was carried out at endpoint. TEX-VAL Michael Castiglione 4 2019-07-11 Rusyn_TAMU 226
View/Edit 1801100004 2018-08-29 TOX-CC SQL-SAL 1.5 Fluidic-3D -No Diseases- 358 0 0 0 0 0 1 MIT SQL-SAL study that observed the effects of 150µM Trovofloxa, 1µg/mL LPS, or the combination of those two, on albumin, LDH, and TNF-alpha production TC2T Michael Castiglione 4 2019-07-10 Cirit_MIT 227
View/Edit 1801100002 2018-07-31 CC SQL-SAL 1.5 Fluidic-3D -No Diseases- 360 0 0 0 0 0 1 MIT study of terfenadine metabolism in SQL-SAL 1.5 model. Three chips were treated with 8.7µM for 10 days. TC2T Michael Castiglione 4 2019-07-10 Cirit_MIT 228
View/Edit RFE1_Variable Matrix Conditions within vLAMPS (PEG, Collagen, Collagen+HGF, Hystem-C, Hystem-HP, ER ECM, PEG-NB) 2018-11-09 CC vLAMPS Fluidic-3D -No Diseases- 253 0 0 0 0 0 0 The purpose of this experiment is to test new matrix conditions with the vLAMPS setting that will aid in matrix stability and hepatocyte function. The conditions to test are PEG, Collagen, Collagen+HGF, Hystem-C, Hystem-HP, ER ECM, PEG-NB. Collagen is the reference control and the stability and functionality will be tested by image analysis and the functional readouts, Albumin, Urea, and LDH. The length of study will be 21 days. UPDDI Dillon Gavlock 4 2019-08-13 Taylor_EPA 229
View/Edit Biomaterial Testing for Overlay 2018-11-07 CC 2D Mono-layer Hepatocytes Static-2D -No Diseases- 36 0 0 0 0 0 9 2D static cultures of 5 biomaterials to potentially replace collagen type 1 in the vLAMPS model UPDDI Larry Vernetti 4 2019-08-13 Taylor_MPS 230
View/Edit Vascularized Tumor-Tier1-3D 2018-07-11 TOX-EFF-CC Vascularized Tumor Model Fluidic-3D -No Diseases- 2,771 0 1,020 0 0 0 1 Microfluidic devices, endothelial cells, cancer cells received from developer’s lab at UC-Irvine for the 3D vascularized tumor model testing, supporting human lung fibroblasts were purchased from Lonza. Drug effects on tumor and vessel network in the 3D vascularized tumor model were evaluated using EPC-AD endothelial cells and HCT116 colon cancer cells. TEX-VAL Michael Castiglione 4 2019-11-13 Rusyn_TAMU 231
View/Edit Tier1a_UCB_3D 2018-05-18 CC Liver Sinusoid Fluidic-3D -No Diseases- 345 0 2 0 0 0 2 This tier of testing was part of the 3D “Baseline” study to measure the secretome of cells seeded in the UC-Berkeley tissue chip without any treatments. In this study, 4 chips were seeded with primary hepatocytes from Lonza, after coating the chips with ECM. Cells were cultured over 11 days with daily media sampling. Media was tested for flow rate, Albumin, Urea-Nitrogen, LDH, and TNF-a. Note: There are no 2D companion studies using the Lonza primary cells, as they were non-adherent in the plates. TEX-VAL Michael Castiglione 4 2019-07-30 Rusyn_TAMU 232
View/Edit Tier1b_UCB_3D 2018-06-09 CC Liver Sinusoid Fluidic-3D -No Diseases- 966 0 0 0 0 0 2 This tier of testing was part of the 3D “Metabolism” study to measure the metabolic capacity of cells seeded in UC-Berkeley tissue chips. In this study, 10 chips were seeded with primary hepatocytes from Lonza, after coating the chips with ECM. Cultures were treated with either a vehicle (0.1% DMSO, n=5), or a 10uM Terfenadine (n=5). Cells were cultured over 13 days with daily media sampling. Media was tested for flow rate, Albumin, Urea-Nitrogen, LDH, and TNF-a, as well as the presence of the parent compound (terfenadine), and the major metabolite (fexofenadine). Additionally, viability testing was carried out at endpoint. Note that the syringe pumps stalled out on day 7. TEX-VAL Michael Castiglione 4 2019-07-30 Rusyn_TAMU 233
View/Edit Tier1c_UCB_3D 2018-07-19 TOX Liver Sinusoid Fluidic-3D -No Diseases- 2,356 0 51 0 0 0 2 This tier of testing was part of the 3D “Toxicity” study to measure the effects of drugs on cells seeded in UC-Berkeley tissue chips. In this study, 30 chips were seeded with primary hepatocytes from Lonza, after coating the chips with ECM. Cultures were treated with either a vehicle (0.1% DMSO, n=6), caffeine (600uM, n=3), trovafloxacin (150uM, n=3), troglitazone (28uM, n=3), tolcapone (88uM, n=3), rosiglitazone (0.8uM, n=3), pioglitazone (3uM, n=3), trovaflocacin + LPS (150uM + 1ug/mL, n=3), or LPS (1ug/mL, n=3). Cells were cultured over 10 days with daily media sampling. Media was tested for flow rate, Albumin, Urea-Nitrogen, LDH, and TNF-a, as well as the presence of the parent drug compounds. Additionally, viability testing was carried out at endpoint. Note: This study was carried out in 2 “arms” of testing: set 1 and set 2 as noted on PTMs. TEX-VAL Michael Castiglione 4 2019-07-30 Rusyn_TAMU 234
View/Edit Tier2a2b_UCB_Liver_3D 2018-08-14 CC Liver Sinusoid Fluidic-3D -No Diseases- 956 0 20 0 0 0 2 This tier of testing was part of the 3D “Baseline” and “Metabolism” studies to measure the secretome and metabolic capacity of cells seeded in UC-Berkeley chips. In this study, 10 chips were seeded with iPSC-derived hepatocytes from CDI after coating the chips with ECM. Cultures were treated with either cell culture media (n=4), vehicle (0.1% DMSO, n=3), or a 10uM Terfenadine (n=3). Cells were cultured over 12 days with daily media sampling. Media was tested for Albumin, Urea-Nitrogen, LDH, and TNF-a, as well as the presence of the parent compound (terfenadine), and the major metabolite (fexofenadine). Additionally, viability testing was carried out at endpoint. Note that there was a power outage that stalled syringe pumps on days 11-12. TEX-VAL Michael Castiglione 4 2019-07-30 Rusyn_TAMU 235
View/Edit Tier2c_UCB_Liver_3D 2018-08-31 TOX Liver Sinusoid Fluidic-3D -No Diseases- 2,138 0 53 0 0 0 2 This tier of testing was part of the 3D “Toxicity” study to measure the effects of drugs on cells seeded in UC-Berkeley chips. In this study, 27 chips were seeded with iPSC-derived hepatocytes from CDI after coating the chips with ECM. Cultures were treated with either a vehicle (0.1% DMSO, n=3), caffeine (600uM, n=3), trovafloxacin (150uM, n=3), troglitazone (28uM, n=3), tolcapone (88uM, n=3), rosiglitazone (0.8uM, n=3), pioglitazone (3uM, n=3), trovaflocacin + LPS (150uM + 1ug/mL, n=3), or LPS (1ug/mL, n=3). Cells were cultured over 10 days with daily media sampling. Media was tested for flow rate, Albumin, Urea-Nitrogen, LDH, and TNF-a, as well as the presence of the parent drug compounds. Additionally, viability testing was carried out at endpoint. TEX-VAL Michael Castiglione 4 2019-07-30 Rusyn_TAMU 236
View/Edit Tier2a2b_UCB_2D 2018-08-14 CC Liver Sinusoid [TAMU 96w] Static-2D -No Diseases- 916 0 60 0 0 0 2 This study is a 2D companion to Tier2a2b_UCB_Liver_3D This tier of testing was part of the 2D “Baseline” and “Metabolism” studies to measure the secretome and metabolic capacity of cells plated in 2D culture. In this study, 30 wells from a 2D cell culture plate (96-well plate) were plated with iPSC-derived hepatocytes from CDI. Cultures were treated with either cell culture media (n=10), vehicle (0.1% DMSO, n=10), or a 10uM Terfenadine (n=10). Cells were cultured over 12 days with daily media sampling. Media was tested for Albumin, Urea-Nitrogen, LDH, and TNF-a, as well as the presence of the parent compound (terfenadine), and the major metabolite (fexofenadine). Additionally, viability testing was carried out at endpoint. TEX-VAL Michael Castiglione 4 2019-07-30 Rusyn_TAMU 239
View/Edit Tier2c_UCB_2D 2018-08-31 TOX Liver Sinusoid [TAMU 96w] Static-2D -No Diseases- 1,946 0 120 0 0 0 2 This study is a 2D companion to Tier2c_UCB_Liver_3D This tier of testing was part of the 2D “Toxicity” study to measure the effects of drugs on cells plated in 2D culture. In this study, 60 wells from a 2D cell culture plate (96-well plate) were plated with iPSC-derived hepatocytes from CDI. Cultures were treated with either a vehicle (0.1% DMSO, n=12), caffeine (600uM, n=6), trovafloxacin (150uM, n=6), troglitazone (28uM, n=6), tolcapone (88uM, n=6), rosiglitazone (0.8uM, n=6), pioglitazone (3uM, n=6), trovaflocacin + LPS (150uM + 1ug/mL, n=6), or LPS (1ug/mL, n=6). Cells were cultured over 10 days with daily media sampling. Media was tested for Albumin, Urea-Nitrogen, LDH, and TNF-a, as well as the presence of the parent drug compounds. Additionally, viability testing was carried out at endpoint. TEX-VAL Michael Castiglione 4 2019-09-03 Rusyn_TAMU 240
View/Edit 1801100005 2018-09-26 TOX SQL-SAL 1.5 Fluidic-3D -No Diseases- 238 0 0 0 0 0 1 Pitt Liver MPS (SQL-SAL) tolcapone and caffeine toxicity TC2T Michael Castiglione 4 2019-07-10 Cirit_MIT 253
View/Edit White Adipose Tier 1 2019-01-01 CC White Adipose Tissue, White Adipose [TAMU 384w] Fluidic-3D, Static-2D -No Diseases- 0 0 8 0 0 0 0 Chip functionality and technology transferability validation using devices provided by developer's lab at UC-Berkeley and 3T3-L1 mouse fibroblasts from ATCC. Experiment in 2D well culture was carried out at the same time for comparison. Undifferentiated 3T3-L1 cells were cultured in differentiation induction medium, loaded into the 3D platform or well plate, cultured for 18 days in chip. Cells were imaged to confirm viability, and maturation to adipocytes. TEX-VAL Admin Sandra Karcher 4 2019-07-11 Rusyn_TAMU 255
View/Edit White Adipose 2D Tier 2 3T3-L1 2018-10-24 TOX White Adipose [TAMU 384w] Static-2D -No Diseases- 2,608 0 10 0 0 0 2 3T3-L1 mouse fibroblasts from ATCC were seeded in 2D well plate and exposed to the same chemical compounds tested in the 3D model as a 2D comparison to the 3D model. TEX-VAL Michael Castiglione 4 2019-08-12 Rusyn_TAMU 260
View/Edit White Adipose 2D Tier 2 HSP 2018-09-11 TOX White Adipose [TAMU 384w] Static-2D -No Diseases- 2,128 0 10 0 0 0 2 Human subcutaneous pre-adipocytes from Zen-Bio were seeded in 2D well plate and exposed to the same chemical compounds tested in the 3D model as a 2D comparison to the 3D model. TEX-VAL Michael Castiglione 4 2019-08-12 Rusyn_TAMU 261
View/Edit White Adipose 3D Tier 2 3T3-L1 2018-10-08 TOX White Adipose Tissue Fluidic-3D -No Diseases- 300 0 10 0 0 0 2 3T3-L1 mouse fibroblasts from ATCC were seeded in the 3D white adipose tissue-on-a-chip platform provided by developer's lab at UC-Berkeley, exposed to selected chemicals for 8 days. At the end of the exposure, LDH, viability, adiponectin secretion were quantified. TEX-VAL Michael Castiglione 4 2019-10-04 Rusyn_TAMU 262
View/Edit White Adipose 3D Tier 2 HSP 2018-09-11 TOX White Adipose Tissue Fluidic-3D -No Diseases- 300 0 10 0 0 0 2 3T3-L1 mouse fibroblasts from ATCC were seeded in the 3D white adipose tissue-on-a-chip platform provided by developer's lab at UC-Berkeley, exposed to selected chemicals for 8 days. At the end of the exposure, LDH, viability, adiponectin secretion were quantified. TEX-VAL Michael Castiglione 4 2019-10-04 Rusyn_TAMU 263
View/Edit RFE3_vLAMPS Training 2019-02-15 CC vLAMPS Fluidic-3D -No Diseases- 63 0 0 0 0 0 1 The purpose of this experiment is to train our new laboratory members on the vLAMPS platform to prepare them for future work on the platform in respective projects. Note: BUN and LDH assays were not analyzed in this study. The levels of BUN and LDH in efflux media collected at 175 and 300 ul/hr are below the sensitivity of the analytical assays. UPDDI Dillon Gavlock 4 2019-08-12 Taylor_MPS 264
View/Edit 1700400001 2017-10-12 CC Intestinal Enteroid [Monolayer TW] Fluidic-3D -No Diseases- 66 0 0 0 0 0 1 Initial TEER testing of membrane formation of JHU Intestinal Enteroid model by TC2T group at MIT Additional description to be entered by MIT TC2T Michael Castiglione 4 2019-07-11 Cirit_MIT 265
View/Edit 1700400002 2017-10-19 CC Intestinal Enteroid [Monolayer TW] Fluidic-3D -No Diseases- 144 0 0 0 0 0 1 Repeat testing of membrane formation TEER of JHU Intestinal Enteroid model by TC2T group at MIT Additional description to be entered by MIT TC2T Michael Castiglione 4 2019-07-10 Cirit_MIT 266
View/Edit 1700400005 2017-10-23 CC Intestinal Enteroid [Monolayer TW] Fluidic-3D -No Diseases- 114 0 0 0 0 0 1 Repeat of study "1700400005" to test membrane formation TEER of JHU Intestinal Enteroid model by TC2T group at MIT Additional description to be entered by MIT TC2T Michael Castiglione 4 2019-07-10 Cirit_MIT 267
View/Edit 1700400007 2017-11-03 CC Intestinal Enteroid [Monolayer TW], Intestinal Enteroid [TAMU Caco-2 TW] Fluidic-3D, Static-3D -No Diseases- 240 0 0 0 0 0 2 A 24-well plate was seeded with primary J2 intestinal enteroid cultures provided by Johns Hopkins. The constructs' membrane permeability was assayed via Transepithelial Electrical Resistance (TEER) TC2T Michael Castiglione 4 2019-12-04 Cirit_MIT 268
View/Edit 1800700007 2018-03-16 TOX Bone Static-3D -No Diseases- 635 0 0 0 0 0 1 TBE by MIT TC2T Michael Castiglione 4 2019-07-10 Cirit_MIT 269
View/Edit Skin MPS Tier 1 2017-11-01 CC Skin MPS Static-3D -No Diseases- 99 0 6 0 0 0 1 TAMU/IBT Tier-1 Human Skin. The overall study plan is a comparison of human skin constructs from the laboratory of Dr. Angela Christiano at Columbia University with the Organisation for Economic Co-operation and Development (OECD) and industry-recognized standard, Epi-Derm commercially available from MatTek Corporation. The model system used for these Tier1 studies is just the tissue prepared by Columbia. The tissues were constructed in the laboratories of Dr. Angela Christiano. Tissue punches were prepared by her laboratory, placed into a gel with the living dermal side in the gel and the air-exposed surface kept dry. The tissues were shipped to the Texas A&M Institute of Biosciences and Technology overnight with the earliest delivery available via Federal Express. The purpose of the Tier1 studies was to 1) demonstrate that viable tissues could be shipped from the tissue developer’s laboratory in New York to the testing center in Houston and that they would arrive alive and suitable for testing and 2) repeat the testing of drugs that had also been tested by the tissue developer’s laboratory as a measure of reproducibility. TEX-VAL Michael Castiglione 4 2019-12-13 Rusyn_TAMU 273
View/Edit Skin MPS Tier 2 2017-12-01 TOX EpiDerm, EpiDermFT, Skin MPS Static-3D -No Diseases- 1,420 0 151 0 0 0 1 TAMU/IBT Tier-2 Human Skin. The overall study plan is a comparison of human skin constructs from the laboratory of Dr. Angela Christiano at Columbia University with the Organisation for Economic Co-operation and Development (OECD) and industry-recognized standard, Epi-Derm commercially available from MatTek Corporation. The model system used for these Tier2 studies is just the tissue prepared by Columbia. The tissues were constructed in the laboratories of Dr. Angela Christiano. Tissue punches were prepared by her laboratory, placed into a gel with the living dermal side in the gel and the air-exposed surface kept dry. The tissues were shipped to the Texas A&M Institute of Biosciences and Technology overnight with the earliest delivery possible via Federal Express. The Tier2 studies are focused on the use of the skin tissues as a model for assessing the irritation or corrosive properties of chemicals. The industry standard has always been an animal model. It has long been desired that an in vitro alternative to the use of animals be found. The OECD has evaluated many models and the EpiDerm model available from MatTek Corporation has been recognized as an alternative. The purpose of the Tier2 studies was to compare the Columbia skin construct with the MatTek EpiDerm and EpiDerm-FT models for their ability to predict 1) whether or not a compound acts as a skin irritant and 2) whether or not a compound acts as a skin corrosive agent. For these studies, six compounds were selected that have known properties and that have been evaluated in many in vitro model systems. The compounds tested were diethyl phthalate (CAS no. 84-66-2) a known non-irritant, 1-bromohexane (CAS no. 111-25-1) a known skin irritant, 1,9-decadiene (CAS no. 1647-16-1) a known non-corrosive compound that can be a skin irritant, isostearic acid (CAS no. 30399-84-9) a known non-corrosive compound, phosphorous tribromide (CAS no. 7789-60-8) a known skin corrosive and hydrochloric acid (CAS no. 7647-01-0) also a known skin corrosive compound. Tissue viability after compound exposure is used by the models to determine whether or not a compound is an irritant or corrosive. The post-assay viability was measured by using PrestoBlue. Three aliquots were removed from each tissue sample and read in a fluorescent plate reader. These RFU values were reported to the database. The mean of the three replicates was then normalized to the water-treated negative controls. These values are reported in the MPD database as ‘% Control’ and are used by the models to predict reactivity. The corrosion test used 8N KOH as a positive control and the irritation test used 5% SDS as a positive control. The predictions of irritant, corrosive or not are reported in the MPS database as either a value of 0 meaning no effect or a value 1 meaning the compound was predicted by the model to be an irritant or corrosive depending on the assay being conducted. TEX-VAL Michael Castiglione 4 2019-12-13 Rusyn_TAMU 275
View/Edit Heart_Tier2_CDI_3D 2018-08-10 TOX Cardiac MPS Fluidic-3D -No Diseases- 3,960 0 495 47 0 0 2 This tier of testing was part of the 3D heart chip study to measure the effects of drugs on cardiomyocytes seeded within the UC-Berkeley chips. In this study, 55 chips were seeded with iPSC-derived cardiomyocytes purchased from CDI (iCell Cardiomyocytes, lot #1293677) in a central growth chamber. Cells were cultured over 4 days in the chip prior to testing with reference compounds. Cultures were treated with either vehicle (n=9, 0.1% DMSO), atenolol (0.01µM, n=4; 0.1µM, n=3; 1µM, n=3; 10µM, n=3), sorafenib (0.1µM, n=4; 1µM, n=3; 10µM, n=3; 100µM, n=3) or carbamazepine (0.1, 1, 10, 100µM, n=5 each) over a 1 week period. At each timepoint (baseline, 5 hours, and daily for days 1-7), phase contrast images were captured (10FPS), and analyzed using the MotionGUI script in Matlab (provided by UC-Berkeley). The outputs of this program were beat rate (BPM), time interval (s, duration between contraction and relaxation), contraction velocity (px/s), and relaxation velocity (px/s). Immunohistochemistry was difficult in this chip, so images are displayed as phase contrast only. It should be noted that while CDI cells performed well for the first few days of treatment, after 3-4 days many chips stopped beating, even the vehicle controls. This was also noted in the 2D side-by-side comparison-- while cells were still viable and fluxing calcium in 2D, they were not physically beating, therefore the motion tracking program did not work for these later days of culture. TEX-VAL Michael Castiglione 4 2019-09-27 Rusyn_TAMU 277
View/Edit Heart_Tier2_CDI_2D 2018-07-14 TOX Cardiac Static [TAMU 384w] Static-2D -No Diseases- 3,464 0 393 41 0 0 2 This tier of testing was a 2D companion to the 3D heart chip study to measure the effects of drugs on cardiomyocytes seeded within the UC-Berkeley chips. In this study, 220 wells were seeded with iPSC-derived cardiomyocytes purchased commercially from CDI into a 384-well cell culture plate. Cells were cultured over 4 days in the plate prior to testing with reference compounds. Cultures were treated with either a media control (n=20), vehicle (0.1% DMSO, n=20), atenolol (0.01, 0.1, 1, 10uM, n=15), sorafenib (0.1, 1, 10, 100uM, n=15), or carbamazepine (0.1, 1, 10, 100uM, n=15) over a week-long exposure, measuring daily. At each timepoint, phase contrast images were captured (10FPS), and analyzed using the MotionGUI script in Matlab (provided by UC-Berkeley). The outputs of this program were beat rate (BPM), time interval (s, duration between contraction and relaxation), contraction velocity (px/s), and relaxation velocity (px/s). As an additional endpoint, cells were treated with a calcium sensitive dye on day 11, and calcium flux was measured as beats/min. Note: Cells stopped physically beating at around day 7 (day 3 of treatment), but calcium flux was still detectable at endpoint on day 11. TEX-VAL Michael Castiglione 4 2019-09-27 Rusyn_TAMU 278
View/Edit TAMU Skeletal Muscle Myobundle Tier 1 2018-06-19 CC Skeletal Myobundle Static-3D -No Diseases- 18,528 0 1,110 1,116 0 0 1 TAMU/IBT Tier-1 Human Skeletal Muscle Myobundles. The overall study plan is an evaluation of the acute and chronic effects of drugs on the ability of skeletal muscle fiber microphysiological systems to respond to electrical stimulation with either a twitch or tetanus response. The tissues were constructed using cells isolated from patients and provided by Dr. George Truskey’s laboratory at Duke University. In addition to the frames, Dr. Truskey’s group provided all of the frames that had ecoflex bands attached. The PDMS plates and molds were made by Dr. Arum Han’s research group at Texas A&M in College Station. The components were assembled, the cells propagated and plated into the devices, and drug treated and tested at the Texas A&M Institute of Biosciences and Technology in Houston. The purpose of the Tier1 studies was to 1) demonstrate that a laboratory other than the where the MPS was developed could successfully make fully functional myobundles that would respond to electrical stimulation and 2) repeat the testing of drugs that had also been tested by the tissue developer’s laboratory as a measure of reproducibility. TEX-VAL Michael Castiglione 4 2019-12-01 Rusyn_TAMU 279
View/Edit 1800600004 2018-03-22 CC Cardiac MPS Fluidic-3D -No Diseases- 24 0 0 0 0 0 1 An initial, technology transfer study for the Berkeley Cardiac Chip to MIT. The study consisted of eight total chips seeded with cardiomyocytes and stromal cells from the Healy Lab at UC Berkeley. It looked the response of 1µM staurosporine on beat rate, as compared to a no compound control. The beat rate was manually calculated from collected video. TC2T Michael Castiglione 4 2019-07-10 Cirit_MIT 282
View/Edit 1800600006 2018-06-06 TOX Cardiac MPS, Cardiac Static [MIT 96w] Fluidic-3D, Static-2D -No Diseases- 28 0 0 0 0 0 1 MIT TC2T eight chip Cardiac MPS study of doxorubicin toxicity. The chips were seeded with with cardiomyocytes and stromal cells from the Healy Lab at UC Berkeley. It looked at the response of 10µM doxorubicin on beat rate, as compared to a no compound control. Beat rate was manually calculated. The chip studies were carried out side-by-side with cardiomyocytes/stromal cells grown in a 96-well static culture format. TC2T Michael Castiglione 4 2019-07-10 Cirit_MIT 283
View/Edit MCF7 Monoculture Plate Study 1 2017-09-09 DM MCF7 Metastatic Breast Cancer Monoculture Model Static-2D -No Diseases- 114 0 72 0 0 0 0 This study was used a growth control for the 3D static and 3D LAMPS model + MCF7 mutant cells. While both mutants (Y537S and D538G) demonstrate increased constitutive growth compared to WT in the absence of estrogen, the D538G muatnt shows enhanced estrogen dependent growth compared to the Y537S mutant. This is in contrast to observations made in co-culture and LAMPS models, where the Y537S mutant shows estrogen-enhanced growth. UPDDI Dillon Gavlock 4 2019-08-13 Taylor_MPS 286
View/Edit MCF7 Monoculture Plate Study 2 2018-03-23 DM MCF7 Metastatic Breast Cancer Monoculture Model Static-2D -No Diseases- 114 0 72 0 0 0 0 monoculture study examining the growth of WT and ESR1 mutants in the absence or presence of estrogen at 20% oxygen tension. While both mutants (Y537S and D538G) demonstrate increased constitutive growth compared to WT in the absence of estrogen, the D538G muatnt shows enhanced estrogen dependent growth compared to the Y537S mutant. This is similar to observations made in monoculture studies performed at 5% oxygen tension, but differs from results obtained in co-culture and LAMPS where the Y537S mutant displays enhanced estrogen dependent growth. UPDDI Dillon Gavlock 4 2019-08-13 Taylor_MPS 287
View/Edit MCF7 Co-Culture Plate Study 2 2018-03-09 DM LAMPS MCF7 Metastatic Breast Cancer Plate Model Static-3D -No Diseases- 114 0 72 0 0 0 0 While both mutants (Y537S and D538G) demonstrate increased constitutive growth compared to WT in the absence of estrogen, the Y537S muatnt shows enhanced estrogen dependent growth compared to the D538G mutant. This is similar to observations made in LAMPSmodels, but different to phenotypes observed in monoculture where the D538G mutant shows estrogen-enhanced growth. UPDDI Dillon Gavlock 4 2019-08-13 Taylor_MPS 288
View/Edit MCF7 Monoculture Plate Study 3 2018-02-16 DM MCF7 Metastatic Breast Cancer Monoculture Model Static-2D -No Diseases- 114 0 72 0 0 0 0 monoculture studies performed at zone 3 (3-6%) oxygen tension examining the growth of WT ESR1 and ESR1 mutants mimicked the results obtained at zone 1(20%) oxygen tension where both mutants displayed constitutive growth increase compared to WT and the D538G mutant showed enhanced estrogen dependent growth. UPDDI Dillon Gavlock 4 2019-08-13 Taylor_MPS 289
View/Edit MCF7 Co-Culture Plate Study 3 2018-03-09 DM LAMPS MCF7 Metastatic Breast Cancer Plate Model Static-3D -No Diseases- 114 0 72 0 0 0 0 The goal was to examine the effects of changes in oxygen tension on growth phenotypes conferred by ESR1 mutant-expressing cells in co-culture models. Whereas monoculture studies performed at zone 3 (3-6%) mimicked the results obtained at zone 1, both co-culture and LAMPS models performed at low oxygen showed distinct differences compared to studies performed at zone 1 oxygen tension. While the constitutive growth advantage for ESR1 mutants remained unaffected, the enhanced E2-dependent growth for the Y537S mutation and the E2-dependent growth for WT expressing cells were lost. Overall, these results demonstrate the importance of oxygen tension in regulating specific growth phenotypes conferred by ESR1 mutants. UPDDI Dillon Gavlock 4 2019-08-13 Taylor_MPS 290
View/Edit Metabolism in SQL-SAL 1.5 2019-03-29 TOX SQL-SAL 1.5 Fluidic-3D -No Diseases- 1,416 0 0 0 0 0 0 The goal of this study was to collect time dependent metabolism of a 6 drug probe cocktail in the SQL-SAl 1.5 device for PK modeling. The collected time points were 2, 4, 8, 24 ,48 , 72, 96 120 and 144 hours. UPDDI Larry Vernetti 4 2019-07-16 Taylor_EPA 294
View/Edit Tier 2 Vascularized Tumor Model Testing 2019-04-01 TOX Vascularized Tumor Model Fluidic-3D -No Diseases- 990 0 540 0 0 0 2 Microfluidic devices, endothelial cells, cancer cells received from developer’s lab at UC-Irvine for the 3D vascularized tumor model testing, supporting human lung fibroblasts were purchased from Lonza. Drug effects on tumor and vessel network in the 3D vascularized tumor model were evaluated using HUVEC-216 endothelial cells and HCT116 colon cancer cells. TEX-VAL Michael Castiglione 4 2019-11-13 Rusyn_TAMU 296
View/Edit Pitt Liver Tier 0 Drug Binding 2019-04-01 CC RED - SQL-SAL 1.5 Static-2D -No Diseases- 98 0 0 0 0 0 0 In this tier of testing, we investigated the binding of test compounds to proteins in the cell culture media. All test compounds (Caffeine, Pioglitazone, Rosiglitazone, Tolcapone, Troglitazone, and Trovafloxacin) were tested in RED (rapid equilibrium dialysis) plates at 1 and 10uM in PBS buffer, Primary cell culture media, and CDI iPSC media. Results are reported as % free fraction. Media Setting Value to Category: 0 => PBS 1 => Primary Cell Culture Media (Tier 1) 2 => iPSC Cell Culture Media (Tier 2) TEX-VAL Michael Castiglione 4 2019-07-26 Rusyn_TAMU 300
View/Edit UCB Liver Tier 0 Drug Binding 2019-04-01 CC RED - Liver Sinusoid Static-2D -No Diseases- 99 0 0 0 0 0 0 In this tier of testing, we investigated the binding of test compounds to proteins in the cell culture media. All test compounds (Caffeine, Pioglitazone, Rosiglitazone, Tolcapone, Troglitazone, and Trovafloxacin) were tested in RED (rapid equilibrium dialysis) plates at 1 and 10uM in PBS buffer, Primary cell culture media, and CDI iPSC media. Results are reported as % free fraction. Media Setting Value to Category: 0 => PBS 1 => Primary Cell Culture Media (Tier 1) 2 => iPSC Cell Culture Media (Tier 2) TEX-VAL Michael Castiglione 4 2019-07-30 Rusyn_TAMU 301
View/Edit 1700600001 2018-06-12 CC Cardiac Static [MIT 96w] Static-2D -No Diseases- 86 0 0 0 0 0 1 MIT experiment with UC Berkeley Cardiac Model. CDI iCell Cardiomyocytes seeded into 96-well plate and treated with isoproterenol. This study was done to develop metrics (Beat Rate, FABP3) used in later studies. TC2T Michael Castiglione 4 2019-07-16 Cirit_MIT 302
View/Edit 1799900001 - Nonspecific Compound Binding in NVU Device 2017-12-17 CC Blood-Brain Barrier (NVU) Fluidic-3D -No Diseases- 160 0 0 0 0 0 2 "This study examined the non-specific binding of five drugs to a cell free NVU device, which is the device used to create the Vanderbilt Blood-Brain Barrier model. The five drugs examined were Bosentan, Dicclofenac, Doxorubicin, Lidocaine, and Fexofenadine. The drugs were added to the cell-free device as a combined cocktail at 1µM. Drug concentrations were measured via mass spectroscopy. Measurements were taken from the cocktail before adding to the device, in addition to three measurements taken from device outflow after 1, 24, and 48 hours in the device." TC2T Michael Castiglione 4 2019-12-04 Cirit_MIT 307
View/Edit 180600007 2018-08-09 TOX Cardiac MPS Fluidic-3D -No Diseases- 30 0 0 0 0 0 1 This study tested the effect of an isoprotenerol and a terfenadine dose escalation on beat frequency in the Healy (UC Berkeley) Heart model. Heart chips were dosed with an increasing amount of compound (0.1µM -> 1.0µM -> 10µM) for 30 minutes per dose. Beat frequency was calculated by manual analysis of video recorded after the 30 minute dose. TC2T Michael Castiglione 4 2019-11-21 Cirit_MIT 311
View/Edit 180600008 2018-08-09 TOX Cardiac MPS Fluidic-3D -No Diseases- 33 0 0 0 0 0 1 This study tested the effect of an verapamil dose escalation and a lidocaine dose escalation on beat frequency in the Healy (UC Berkeley) Heart model. Heart chips were dosed with an increasing amount of compound (0.01µM->0.1µM->1.0µM for verapamil; 0.1µM -> 1.0µM -> 10µM for lidocaine) for 30 minutes per dose. Beat frequency was calculated by manual analysis of video recorded after each 30 minute dose. TC2T Michael Castiglione 4 2019-11-21 Cirit_MIT 312
View/Edit 180600009 2018-08-15 TOX Cardiac MPS Fluidic-3D -No Diseases- 64 0 0 0 0 0 1 This study tested the effect of an lidocaine dose escalation and a doxorubicin dose escalation on beat frequency in the Healy (UC Berkeley) Heart model. Heart chips were dosed with an increasing amount of compound (1µM->10µM->100µM) for 30 minutes per dose. Beat frequency was calculated by manual analysis of video recorded after each 30 minute dose. The doxorubicin treated chips continued the treatment at 100µM for 2 days after the dose escalation was complete TC2T Michael Castiglione 4 2019-11-21 Cirit_MIT 313
View/Edit 1899900002 - Nonspecific Compound Binding in Bone Tumor Device 2018-12-17 CC Bone Static-3D -No Diseases- 60 0 0 0 0 0 2 "This study examined the non-specific binding of five drugs to HeLiVaSkCa - Cancer Module device produced at Columbia. This device was used in the Bone Tumor model. The five drugs examined were Bosentan, Dicclofenac, Doxorubicin, Lidocaine, and Fexofenadine. The drugs were added to the cell-free device as a combined cocktail at 1µM. Drug concentrations were measured via mass spectroscopy. Measurements were taken from the cocktail before adding to the device, and after 48 hours in the device." TC2T Michael Castiglione 4 2019-12-04 Cirit_MIT 316
View/Edit 1799900001 - Nonspecific Compound Binding in Brain Device Study 1 2017-12-17 CC Brain [MIT Cell Free] Static-3D -No Diseases- 140 0 0 0 0 0 2 "This study examined the non-specific binding of five drugs to a cell free CN Bio T18 platform, both with PEG gel + transwell insert and with the bare platform. This platform is used in the UW Thomson Brain model. The five drugs examined were Bosentan, Dicclofenac, Doxorubicin, Lidocaine, and Fexofenadine. The drugs were added to the cell-free device as a combined cocktail at 1µM. Drug concentrations were measured via mass spectroscopy. Measurements were taken from the cocktail before adding to the device, and after 48 hours in the device." TC2T Michael Castiglione 4 2019-12-04 Cirit_MIT 317
View/Edit 1899900002 - Nonspecific Compound Binding in Brain Device Study 2 2018-12-17 CC Brain Static-3D -No Diseases- 75 0 0 0 0 0 2 "This study examined the non-specific binding of five drugs to a cell free Costar 24-well transwell plate with PEG gel. This platform was used in the UW Thomson Brain model. The five drugs examined were Bosentan, Dicclofenac, Doxorubicin, Lidocaine, and Fexofenadine. The drugs were added to the cell-free device as a combined cocktail at 1µM. Drug concentrations were measured via mass spectroscopy. Measurements were taken from the cocktail before adding to the device, and after 48 hours in the device." TC2T Michael Castiglione 4 2019-12-04 Cirit_MIT 318
View/Edit 1899900002 - Nonspecific Compound Binding in Cardiac Device 2018-12-17 CC Cardiac MPS Fluidic-3D -No Diseases- 60 0 0 0 0 0 2 This study examined the non-specific binding of five drugs to Cardiac MPS scaffold device produced at UC Berkeley. This device was used in the Healy Cardiac model. The five drugs examined were Bosentan, Dicclofenac, Doxorubicin, Lidocaine, and Fexofenadine. The drugs were added to the cell-free device as a combined cocktail at 1µM. Drug concentrations were measured via mass spectroscopy. Measurements were taken from the cocktail before adding to the device, and after 48 hours in the device. TC2T Michael Castiglione 4 2019-12-04 Cirit_MIT 319
View/Edit 1899900002 - Nonspecific Compound Binding in Intestinal Enteroid Device 2018-12-17 CC Intestinal Enteroid [Monolayer TW] Fluidic-3D -No Diseases- 75 0 0 0 0 0 2 This study examined the non-specific binding of five drugs to a Costar 24 transwell plate. This plate was used in the Donowitz Intestinal Enteroid model. The five drugs examined were Bosentan, Dicclofenac, Doxorubicin, Lidocaine, and Fexofenadine. The drugs were added to the cell-free device as a combined cocktail at 1µM. Drug concentrations were measured via mass spectroscopy. Measurements were taken from the cocktail before adding to the device, and after 48 hours in the device. TC2T Michael Castiglione 4 2019-12-04 Cirit_MIT 320
View/Edit 1799900001 - Nonspecific Compound Binding in Kidney Proximal Tubule Device 2017-12-17 CC Kidney Proximal Tubule [Fluidic] Fluidic-3D -No Diseases- 150 0 0 0 0 0 2 This study examined the non-specific binding of five drugs to a cell free Nortis single channel device. This device was used in the UWash Kidney Promixal Tubule model. The five drugs examined were Bosentan, Dicclofenac, Doxorubicin, Lidocaine, and Fexofenadine. The drugs were added to the cell-free device as a combined cocktail at 1µM. Drug concentrations were measured via mass spectroscopy. Measurements were taken from the cocktail before adding to the device, and after 3, 24, and 48 hours in the device. TC2T Michael Castiglione 4 2019-12-04 Cirit_MIT 321
View/Edit 1899900002 - Nonspecific Compound Binding in MiniBrain Device 2018-12-17 CC Brain Spheroids Fluidic-3D -No Diseases- 60 0 0 0 0 0 2 This study examined the non-specific binding of five drugs to a 6 well plate. This plate was used in the JHU Brain Spheroid/Mini-Brain model. The five drugs examined were Bosentan, Dicclofenac, Doxorubicin, Lidocaine, and Fexofenadine. The drugs were added to the cell-free device as a combined cocktail at 1µM. Drug concentrations were measured via mass spectroscopy. Measurements were taken from the cocktail before adding to the device, and after 48 hours in the device. TC2T Michael Castiglione 4 2019-12-04 Cirit_MIT 322
View/Edit 1899900002 - Nonspecific Compound Binding in Skin Device 2018-12-17 CC Skin MPS Static-3D -No Diseases- 60 0 0 0 0 0 2 This study examined the non-specific binding of five drugs to the Columbia HeLiVaSkCa Skin Module device. This device was used in the Columbia Skin model. The five drugs examined were Bosentan, Diclofenac, Doxorubicin, Lidocaine, and Fexofenadine. The drugs were added to the cell-free device as a combined cocktail at 1µM. Drug concentrations were measured via mass spectroscopy. Measurements were taken from the cocktail before adding to the device, and after 48 hours in the device. TC2T Michael Castiglione 4 2019-12-04 Cirit_MIT 323
View/Edit 1801100005 - Nonspecific Compound Binding in SQL-SAL Nortis Device 2018-10-01 CC SQL-SAL 1.5 Fluidic-3D -No Diseases- 111 0 0 0 0 0 2 This study examined the non-specific binding of three drugs to a cell free Nortis single channel device. This device was used in the Pittsburgh SQL-SAL model. Note that the device is the same Nortis device used in the Kidney Proximal Tubule model, and it has had non-specific binding for 5 additional drugs tested in study "1799900001 - Non-Specific Compound Binding in Kidney Device" The three drugs examined were Terfenadine (10µM), Cerivastatin (1µM), and Diclofenac (1µM). Drug concentrations were measured via mass spectroscopy. Measurements were taken from the stock before adding to the device, and daily after 0 to 10 day in the device. TC2T Michael Castiglione 4 2019-12-04 Cirit_MIT 324
View/Edit Heart_Tier1_UCB_2D_a 2019-05-30 TOX-CC Cardiac Static [TAMU 384w] Static-2D -No Diseases- 1,248 0 96 20 0 0 1 This study is a repeat of “Heart_Tier1_UCB_2D”. In the original study, only phase contrast video capture was performed, due to issues with the calcium-sensitive dye. In this study, the same treaments/exposures were repeated, capturing both the phase contrast videos for motion analysis, as well as the calcium flux data. This tier of testing was a 2D companion to the 3D heart chip study to measure the effects of drugs on cardiomyocytes seeded within the UC-Berkeley chips. In this study, 160 wells were seeded with iPSC-derived cardiomyocytes provided by UC-Berkeley, as well as supporting fibroblasts (iPS-stromal cells) in a 384-well cell culture plate. Cells were cultured over 4 days in the plate prior to testing with reference compounds. Cultures were treated with either a media control, vehicle (0.1% DMSO), cisapride, verapamil, or isoproterenol for a 25 minute exposure at concentrations of 1, 10, 100, 1000, and 10000nM (n=10 for all controls and treatments). After each exposure, phase contrast images were captured (10FPS), and analyzed using the MotionGUI script in Matlab (provided by UC-Berkeley). The outputs of this program were beat rate (BPM), time interval (s, duration between contraction and relaxation), contraction velocity (px/s), and relaxation velocity (px/s). Additionally, a calcium sensitive dye was added to capture calcium flux at baseline and after treatment. TEX-VAL Courtney Sakolish 4 2019-10-01 Rusyn_TAMU 336
View/Edit TAMU Skeletal Muscle Myobundle Tier 2 2018-06-19 TOX Skeletal Myobundle Static-3D -No Diseases- 3,040 0 1,504 1,504 0 0 0 TAMU/IBT Tier-2 Human Skeletal Muscle Myobundles. The overall study plan is an evaluation of the acute and chronic effects of drugs on the ability of skeletal muscle fiber microphysiological systems to respond to electrical stimulation with either a twitch or tetanus response. The tissues were constructed using cells isolated from patients and provided by Dr. George Truskey’s laboratory at Duke University. In addition to the frames, Dr. Truskey’s group provided all of the frames that had ecoflex bands attached. The PDMS plates and molds were made by Dr. Arum Han’s research group at Texas A&M in College Station. The components were assembled, the cells propagated and plated into the devices, and drug treated and tested at the Texas A&M Institute of Biosciences and Technology in Houston. The purpose of the Tier2 studies was to extend the testing of the MPS beyond that of the previous work by the tissue developer. In this instance the TAMU/IBT testing center tested four additional drugs for acute and chronic effects. The drugs were Sunitinib, Nelarabine, Metformin, and Erlotinib. These drugs are suspected of causing rhabdomyolysis a side effect of drug treatment resulting in extreme muscle fatigue or nonresponsiveness. TEX-VAL Michael Castiglione 4 2019-12-13 Rusyn_TAMU 341
View/Edit PBPK-Continuous Infusion-Metabolism in LAMPS 2019-05-29 PK LAMPS Fluidic-3D -No Diseases- 192 0 0 0 0 0 0 The goal of this study was to collect time dependent metabolism of a 6 drug probe cocktail in the LAMPS device for PK modeling. The collected time points were 2, 4, 8, 24 ,48 , 72, 96 120 and 144 hours. UPDDI Tongying Shun 4 2020-05-21 Taylor_MPS 400
View/Edit PBPK-Continuous Infusion-Metabolism in SQL 1.5 2019-03-30 PK SQL-SAL 1.5 Fluidic-3D -No Diseases- 180 0 0 0 0 0 0 Cloned from Study UPDDI-PK-2019-05-29-PBPK-Continuous Infusion-Metabolism in LAMPS. The goal of this study was to collect time dependent metabolism of a 6 drug probe cocktail in the LAMPS device for PK modeling. The collected time points were 2, 4, 8, 24 ,48 , 72, 96 120 and 144 hours. UPDDI Tongying Shun 4 2020-05-21 Taylor_MPS 402
View/Edit PBPK Bolus-Suspension Metabolism_2019-06-15 2019-06-15 PK Hepatocyte Suspension Fluidic-3D -No Diseases- 72 0 0 0 0 0 0 Purpose: the goal of this experiment was to collect 0, 10, 30, 60, 120 and 240 minute data on the elimination of 6 probe compounds and formation of 6 Cyp isoenzyme specific metabolites in suspension culture of primary human hepatocytes for PBPK model development. Methods: Hu1838 lot of primary human hepatocytes were suspended in HMM media containing the 6 probes compounds (Table 1, protocol). Parent and metabolite compounds were extracted from the suspension cultures by acetonitrile. The samples were submitted for mass spec determination. Results: The T1/2 of each parent probe was used to calculate apparent intrinsic clearance for in vitro in vivo extrapolation. UPDDI Tongying Shun 4 2020-05-21 Taylor_MPS 403
View/Edit PBPK Bolus-Suspension Metabolism_2018-06-26 2018-06-26 PK Hepatocyte Suspension Fluidic-3D -No Diseases- 72 0 0 0 0 0 0 Purpose: the goal of this experiment was to collect 0, 10, 30, 60, 120 and 240 minute data on the elimination of 6 probe compounds and formation of 6 Cyp isoenzyme specific metabolites in suspension culture of primary human hepatocytes for PBPK model development. Methods: Hu1838 lot of primary human hepatocytes were suspended in HMM media containing the 6 probes compounds (Table 1, protocol). Parent and metabolite compounds were extracted from the suspension cultures by acetonitrile. The samples were submitted for mass spec determination. Results: The T1/2 of each parent probe was used to calculate apparent intrinsic clearance for in vitro in vivo extrapolation. UPDDI Tongying Shun 4 2020-05-21 Taylor_MPS 404
View/Edit Kidney Study 8 - TERT/RPTEC Characterization - Mimetas Plate 2020-10-29 CC Proximal Tubule/Vascular (Mimetas 3-lane) Fluidic-3D -No Diseases- 332 0 658 12 0 0 4 This study consists of 80 chips across 2 mimetas plates. Four sources of RPTECs were seeded in the tubular channel (RPTEC/TERT1, RPTEC/TERT1-OAT1, RPTEC/TERT1-OCT2, or RPTEC/TERT1-OAT3; 20 chips each), and HUVECs were seeded in the vascular channel of all chips. After injection of cells, plates were perfused via rocking for 8 days, with endpoints including phase contrast imaging, immunocytochemistry, FITC-BSA permeability, LDH release, and pAH (para-aminohippuric acid) secretion. TEX-VAL Courtney Sakolish 4 2022-05-13 TexVal_Consortium 470
View/Edit Tier2and2.1_Lonza_TempOseq 2017-02-27 TOX-OMICS Kidney Proximal Tubule [384-well Static], Kidney Proximal Tubule [Fluidic] Fluidic-3D, Static-2D -No Diseases- 0 510,496 0 0 0 0 0 Tier 2 Kidney Promixal Tubule compound toxicity study with assays of TempOseq transcriptomics. In this study, 17 fluidic chips were perfused with the test compound until significant disruption of the tubule was observed. This study also includes 29 static wells from a 384 well plate, seeded with Lonza renal proximal tubule cells (RPTECs), in which cells are treated with nephrotoxic compounds (cisplain, gentamicin, and cadmium) to study their effects on a monolayer. TEX-VAL Tongying Shun 4 2022-08-31 Rusyn_TAMU 481
View/Edit Tier1_Lonza & KIM_TempOseq 2017-01-27 EFF-OMICS Kidney Proximal Tubule [384-well Static], Kidney Proximal Tubule [Fluidic] Fluidic-3D, Static-2D -No Diseases- 0 870,175 0 0 0 0 0 This TempOseq genetic study includes 17 fluidic MPS chips and 37 static wells (384-well plate) seeded with Lonza renal proximal tubule (RPTEC) cells, and 6 fluidic MPS chips and 20 static wells (384-well plate) seeded with HIM-31 cells from the University of Washington. TEX-VAL Tongying Shun 4 2022-09-01 Rusyn_TAMU 482
View/Edit Kidney Study 1 - 2D Cell characterization 2020-03-20 TOX 2D Cell Culture - Kidney Static-2D -No Diseases- 660 0 192 0 0 0 1 This was a 2D study where RPTECs (Lonza), podocytes (Celprogen), and glomeruluar endothelial cells (Celprogen) were cultured in a 96 well plate and exposed to 10,100, and 1000uM Cisplatin, or 10, 100, and 1000uM Sorafenib for 48h. Wells were phase contrast imaged at 24 and 48h, and effluent was collected for multiplex ELISA to determine the presence or absence of: Calbindin, KIM-1, Renin, TFF3, GST-a, Osteoactivin, TIMP-1, IP-10, FABP-1, and Collagen IV. TEX-VAL Courtney Sakolish 4 2022-05-13 TexVal_Consortium 495
View/Edit Kidney Study 2 - Mimetas Practice Plate 2020-03-24 CC Proximal Tubule/Vascular (Mimetas 3-lane) Fluidic-3D -No Diseases- 30 0 40 0 0 0 2 In this study, 2 operators seeded 20 chips each on a Mimetas OrganoPlate 3-lane (4003-400-B) plate. Here, we practiced gel loading, and cell seeding with Lonza RPTECs and HUVECs, and imaged the tubule formation at 5 days and 14 days of rocking perfusion. Additionally, effluent was collected from a few chips and tested for kidney injury markers via a Multiplex ELISA. TEX-VAL Courtney Sakolish 4 2022-05-13 TexVal_Consortium 496
View/Edit Kidney Study 3 - Proximal tubule/glomerulus test seeding 2020-06-08 TOX-CC Mimetas 3-lane Organoplate Fluidic-3D -No Diseases- 18 0 142 0 0 0 1 In this study, RPTECs (top channel) and HUVECs (bottom channel) were seeded into the left half of the mimetas plate, and glomerular endothelial cells and podocytes were seeded into the right half of the mimetas plate (both cell types in top channel, bottom channel left empty). Cells were cultured under rocking perfusion for a total of 14 days. Treatments (indicated in study design image) were added on day 12 of culture, and cells were exposed for 48h through the HUVEC channel or glomerular channel. Endpoints for this study include phase contrast imaging, permeability studies with FITC-dextran (IMAGING ONLY), and Immunocytochemistry. TEX-VAL Courtney Sakolish 4 2022-05-13 TexVal_Consortium 498
View/Edit Kidney Study 4 - Proximal Tubule/Glomerulus Test plates (Altered ECM) 2020-07-15 TOX-CC Mimetas 3-lane Organoplate Fluidic-3D -No Diseases- 97 0 640 0 0 0 2 In this study, we had 2 plates, each seeded on one half with RPTECs/HUVECs, and on the other half, glomerular endothelial and podocyte cells. Cells were seeded in a staggered fashion, with RPTECS/HUVECs running for 14 days (Day 0-14) under rocking perfusion, and glomerular cells for 9 days under perfusion (days 5-14). Additionally, the ECM of the glomerular chips was increased from 4 to 6mg/mL collagen I + genipin crosslinker to try and avoid issues with invasion that were previously noted. Starting on day 12, chips were treated with compounds for 48h as noted in the attached design image. Chips were phase contrast imaged, tested for permeability with TRITC-BSA, and effluent was collected for LDH testing. TEX-VAL Courtney Sakolish 4 2022-05-13 TexVal_Consortium 499
View/Edit Kidney Study 5 - Mimetas Proximal Tubule (Fresh vs. Commercial RPTECs) 2020-08-14 CC Proximal Tubule/Vascular (Mimetas 3-lane) Fluidic-3D -No Diseases- 152 0 288 0 0 0 3 In this study, we seeded 1 mimetas Organo3-lane plate (#4004) with Freshly isolated primary kidney cells (10 chips), or Lonza commercial primary RPTECs (30 chips). All chips were seeded with the same HUVECs in the endothelial channel. This experiment was carried out over 12 days of rocking perfusion, and treatments started on day 10 and ran 48h through to day 12. Treatments included 1mM pAH (para-aminohippuric acid) +/- 1mM probenecid, and Cisplatin (1, 10, 100, 1000uM). Media was collected and tested for pAH secretion, and LDH release. Phase contrast images were collected at initiation of treatment (d10), and 24 (d11) and 48h (d12) after treatment. At endpoint, viability was determined with L/D Staining. TEX-VAL Courtney Sakolish 4 2022-05-13 TexVal_Consortium 500
View/Edit Kidney Study 6 - Glomerulus (Old vs New Plates) 2020-09-22 TOX-CC Glomerulus (Mimetas 3-lane) Fluidic-3D -No Diseases- 137 0 714 0 0 0 2 In this study, 2 mimetas 3-lane plates were seeded, an old design (cat# 4003) with a smaller opening for seeding, and a new design (cat# 4004) with a larger opening for seeding. In both plates, podocytes and glomerular endothelial cells were layered in a single channel, leaving the second channel free of cells. Half of each plate was injected with the recommended ECM (4mg/mL collagen 1), and the other half was seeded with a higher concentration of collagen (6mg/mL) plus genipin crosslinker to determine effects of ECM stiffness on cell seeding and proliferation. Chips were tested for permeability (FITC-HSA), then fixed on days 3, 6, or 8 across the plates. Additionally, treatments of 0.1 or 1uM Doxorubicin were started on day 6 for 48h until day 8 at endpoint. Unfortunately, in this study, fixing part of the plate with formalin affected other chips that were tested at later timepoints (on days 6 and 8). TEX-VAL Courtney Sakolish 4 2022-05-13 TexVal_Consortium 501
View/Edit Kidney Study 7 - Glomerulus Model Establishment over Time (Day 3 v. Day 9) 2020-10-12 TOX-CC Glomerulus (Mimetas 3-lane) Fluidic-3D -No Diseases- 320 0 696 2 0 0 5 In this study, 2 plates were prepared with podocytes and glomerular endothelial cells to form the glomerulus model. One of the 2 plates was stopped on day 3 of rocking, and the second plate on day 9 of rocking perfusion. In the day 3 plate, a 48 treatment with either vehicle (0.1% DMSO), or 1uM Doxorubicin ran from day 1-3. In the day 9 plate, treatment ran from days 7-9. The purpose of this study was to monitor the formation of the channels over time. Endpoints include daily phase contrast imaging, LDH, podocalyxin, synaptopodin, and nephrin secretion, Permeability (FITC-BSA), and endpoint immunocytochemistry. TEX-VAL Courtney Sakolish 4 2022-05-13 TexVal_Consortium 503
View/Edit Kidney Study 9 - Glomerulus -- Separate seeding of podocytes and glomerular endothelial cells 2020-11-16 TOX-CC Glomerulus (Mimetas 3-lane) Fluidic-3D -No Diseases- 148 0 568 0 0 0 0 In this study, 2 plates (organo 3-lane, #4003) were seeded with podocytes and glomerular endothelial cells. In this study, we wanted to test cell viability and tubule formation when these two cell types were separately seeded, so glomerular endothelial cells were seeded into the top channel, and podocytes into the bottom. 24h after cell injection, perfusion was initiated via rocking, and carried out over 7 days. Cells were treated through the glomerular endothelial channel with either vehicle (0.1% DMSO), Doxorubicin (30uM) or Puromycin (30uM) for 48h between days 5 and 7. At endpoint, channels were tested for permeability (FITC-BSA or FITC-Dextran-150kDa), Viability (live-dead imaging), and LDH activity (effluent testing). TEX-VAL Courtney Sakolish 4 2022-05-13 TexVal_Consortium 504
View/Edit Kidney Study 10 - 2D TERT1 Characterization (Uptake Study) 2020-12-04 CC Kidney Proximal Tubule [96-well Static] Static-2D -No Diseases- 192 0 192 0 0 0 1 In this study, TERT1/RPTEC (basal, OAT1, OCT2, OAT3) cell lines were seeded into a 96 well plate to characterize EAM-1 (OCT2) and 6-CF (OAT1, OAT3) uptake in 2D. Cells were cultured in the plate for 3 days, then uptake assays were carried out (image based, effluent based) in the presence or absence of inhibitors (probenecid, novobiocin, or cimetidine). TEX-VAL Courtney Sakolish 4 2022-05-13 TexVal_Consortium 505
View/Edit Kidney Study 11 - 3D TERT1 Characterization (Uptake Study) 2020-12-14 CC Proximal Tubule/Vascular (Mimetas 3-lane) Fluidic-3D -No Diseases- 92 0 187 0 0 0 3 In this study, a mimetas 3-lane organoplate (#4004) was seeded with TERT1/RPTEC (basal, OAT1, OCT2) cell lines in the top cell channel, and HUVECs in the bottom channel. The ECM was 4mg/mL collagen I in the center lane. Cells were seeded on d0, and endpoint was on d4. To test pAH secretion, on day 2 of culture, 1mM pAH +/- 1mM probenecid was added to the HUVEC channel and incubated from d2-4 (48h treatment). On d4, effluent was collected from both the HUVEC and RPTEC channels and pAH concentration was quantified. To test 6-CF and EAM-1 Uptake, on d3 of culture, other wells were treated with or without inhibitors (100uM probenecid or 100uM cimetidine) for 24h prior to the uptake assay with 6-CF and EAM-1. On d4, RPTEC channels were then treated with 6-CF (3uM) or EAM-1 (5uM) for 20 minutes, then rinsed 4X with ice-cold PBS, and imaged, then lysed for image-based and lysate analysis of internalized fluorophore. TEX-VAL Courtney Sakolish 4 2022-05-13 TexVal_Consortium 506
View/Edit Kidney Study 12 - Glomerulus (3D) - New Glomerular Endothelial Cells 2021-01-26 CC Glomerulus (Mimetas 3-lane) Fluidic-3D -No Diseases- 102 0 232 0 0 0 0 In this study, 1 mimetas plate (#4003) was first loaded with 4mg/mL collagen I in the ECM lane. Primary human podocytes (Celprogen) were then loaded into the top chamber, and allowed 5 hours to attach with the plate on its side. After podocyte attachment, primary human glomerular endothelial cells (Cell Systems) were loaded into the top channel, and layered on top of the podocytes against the gel. The plate was incubated overnight on its side to allow for maximum attachment of the cells to the gel layer. Perfusion was initiated the following morning (d1) by rocking. On d2, media was collected and exchanged with either vehicle (0.1% DMSO), or 30uM puromycin treated media, and incubated for 24h while rocking. On d3, media was collected, and wells were either treated with FITC-BSA (BSA permeability), FITC-dextran 150kDA (dextran permeability), or 4% PFA (fixed for ICC). TEX-VAL Courtney Sakolish 4 2023-05-23 TexVal_Consortium 507
View/Edit RFE91_MAFLD Drug Testing 2021-01-12 EFF-DM LAMPS Fluidic-3D -No Diseases- 637 0 0 0 0 0 2 Goal: Based on the computational pipeline and analysis performed, the goal is to test 3 of the highest-ranking drugs predicted by this analysis in the LAMPS model to determine if treatment with these compounds results in the amelioration of any of the disease phenotypes associated with EMS medium treatment. **Please note that AT7867 treatment resulted in cell toxicity Compounds tested: Troglitazone (10 uM) Pioglitazone (30 uM) AT7867 (10 uM) Obeticholic acid (10 uM; (+) control) UPDDI Mark Miedel 4 2023-08-21 Taylor_MPS 517
View/Edit RFE95_MAFLD Drug Testing 2021-02-08 EFF-DM LAMPS Fluidic-3D -No Diseases- 573 0 0 0 0 0 2 Goal: Based on the computational pipeline and analysis performed, the goal is to test 3 of the highest-ranking drugs predicted by this analysis in the LAMPS model to determine if treatment with these compounds results in the amelioration of any of the disease phenotypes associated with EMS medium treatment. **Please note that AT7867 and A-443654 treatment resulted in cell toxicity Compounds tested: Troglitazone (10 uM) Pioglitazone (30 uM) AT7867 (1 uM) A-443654 (1 uM) UPDDI Mark Miedel 4 2023-08-21 Taylor_MPS 518
View/Edit Lung Study 1 - Model Establishment 2020-02-03 CC Small Airway - ALI (UPenn) Static-3D -No Diseases- 192 0 12 0 0 0 0 In this study, 6 chips were seeded with HMVEC-L (human microvascular endothelial cells from lung; bottom chamber) and SAEC (small airway epithelial cells; top chamber). These chips were airlifted after 24h of seeding, and media was refreshed daily until endpoint at 15 days. On day 15, one chip was treated with fluorescent microbeads to measure ciliary beating, and other were fixed and stained for ZO-1 and nuclei. Media was collected from all chips, pooled, and tested for LDH and mucin secretion. TEX-VAL Courtney Sakolish 4 2022-05-13 TexVal_Consortium 543
View/Edit Liver_Mimetas 2-lane_Protocol optimization 1 2021-05-03 CC Mimetas 2-lane Liver Chip Fluidic-3D -No Diseases- 48 0 1,728 0 0 0 0 In this study, on Day 0, iPS human hepatocytes (iHeps) with collagen were injected in the ECM lane of the Mimetas 2-lane plate. After that, iHep plating media was added to the perfusion channel. Perfusion was initiated immediately by rocking. On Day 5, HMEC-1 cells were loaded into the perfusion channel, and allowed 1 hour to attach with the plate on its side. Then, iHep maintenance media or iHep/HMEC-1 mixed media was added to the perfusion channel. Media was collected and refreshed everyday. The plate was incubated at 37°C with perfusion by rocking. No test articles were treated. On Day 21, half the wells were treated with Calcein/Hoechst/Propidum Iodide (for cell viability) and 4% PFA (fixed for ICC). The assays for albumin, BUN, LDH, and AFP were conducted with collected media. TEX-VAL Yuki Kato 4 2023-05-23 TexVal_Consortium 545
View/Edit RFE108_LAMPS MAFLD drug testing 3 2021-04-26 EFF-DM LAMPS Fluidic-3D NAFLD 675 0 0 0 0 0 1 Goal: Based on the computational pipeline and analysis performed, the goal is to test 2 or 3 of the highest-ranking drugs predicted by this analysis in the LAMPS model to determine if treatment with these compounds results in the amelioration of any of the disease phenotypes associated with EMS medium treatment. Compounds tested: Vorinostat (SAHA) (1.7 uM) Vorinostat (SAHA) (5 uM) AS601245 (1 uM) AS601245 (3 uM) UPDDI Mark Miedel 4 2023-08-21 Taylor_MPS 547
View/Edit CNBio_Proximal Tubule_Exp1 2021-05-24 CC Proximal Tubule (CNBio - T12), Proximal Tubule (Transwell) Fluidic-3D, Static-3D -No Diseases- 240 0 22 0 0 0 0 In this study, TERT1/RPTEC-OAT1 (ATCC) cells were co-cultured with HUVECs (Lonza) in both the CNBio System (TC12) and Transwell cultures (TW). HUVECs were seeded on the basolateral side of the membranes, and TERT1/RPTEC-OAT1 cells were seeded on the apical surface of the membranes. 12 wells/chips were seeded into each growth platform (TC12, TW). 6 wells/chips from each platform were cultured in ATCC TERT1/RPTEC growth media only, and the other 6 were grown in ATCC TERT1/RPTEC growth media supplemented with Lonza EGM-2 supplements to help support the growth of the HUVECs. TC12 cultures were grown under a flow rate of 1uL/s, and TW were static. Cultures were taken out over 6 days with TEER measured throughout, and media collected from the top and bottom for LDH. At endpoint, viability was measured using PrestoBlue, and permeability was measured through the addition of FITC-dextran-150kDa to the basolateral chamber for 1h (and measuring permeability to the apical chamber). Lastly, membranes were fixed and imaged for ZO-1, VE-cadherin, OAT1, and DNA. Conclusions: • TEER was higher overall (by ~10 Ohm*cm2) in static Transwell cultures, compared to their fluidic CNBio counterparts. • TEER began to decrease around day 3 of culture, but was maintained longer in cultures supplemented with the additional HUVEC media supplements compared to the REGM only. • Permeability to FITC-dextran (150kDa) was similar in all conditions, future studies will use a smaller size dextran to parse out differences in permeability. • Immunocytochemistry revealed that by day 6 of culture, there was a loss in epithelial barrier, and the RPTECs began to lift off the membrane. Therefore we should perform endpoint studies prior to observing this dip in TEER over time. • LDH release was low in 3D cultures of cells in the REGM media only, but increased 2-3 fold when cells were grown in static conditions, or when the HUVEC supplements were added to the “coculture” media. • Co-culture of HUVECs and RPTECs was challenging due to different media requirements, and even with HUVEC supplements added into the RPTEC media, viability remained low. Therefore future studies will focus only on monoculture of RPTECs. • HUVECs were grown on the bottom of the Transwell (in the path of fluid flow in the CNBio plate), however we are most interested in the effects of shear on the RPTECs, therefore in future experiments we will try top vs bottom seeding of RPTECs. TEX-VAL Courtney Sakolish 4 2023-05-23 TexVal_Consortium 550
View/Edit CNBio_Proximal Tubule_Exp2 2021-06-03 CC Proximal Tubule (CNBio - T12), Proximal Tubule (Transwell) Fluidic-3D, Static-3D -No Diseases- 348 0 48 0 0 0 0 In this study, TERT1/RPTEC-basal (ATCC) cells were seeded in both the CNBio System (TC12) and Transwell cultures (TW). RPTECs were seeded on either the apical or basolateral surface of the membranes to determine the effects of direct flow on the monocultures. 12 wells/chips were seeded into each growth platform (TC12, TW). 6 wells/chips from each platform were cultured in each seeding configuration (apical or basolateral). TC12 cultures were grown under a flow rate of 1uL/s, and TW were static. Cultures were taken out over 11 days, with media changes and TEER measurements every 48h. At endpoint, cultures were tested for pAH (para-aminohippuric acid) secretion, permeability testing, and viability testing. Conclusions: • Water transport was observed in all culture conditions (static, fluidic; top, bottom seeded) via measured changes in volume (determined by effluent mass) from the top and bottom chambers of wells. The highest degree of water transport was observed when RPTECs were basolaterally seeded, and in fluidic conditions. • Media Absorbance (ABS) was also used to monitor water transport, and was determined to be a less invasive/destructive method for measurement than total effluent mass, as we could just measure a sample of media for ABS rather than the entire liquid volume in the chamber, which caused additional stress on the cells. • LDH was slightly elevated in fluidic cultures compared to static counterparts. There was no observable difference in top vs bottom seeded cultures. • pAH secretion was higher in cells that were seeded on the bottom of the Transwells (vs top seeded), and was also slightly higher in static vs fluidic cultures. • Permeability to FITC-dextran (150kDa) was similar in all conditions (static vs fluidic, top vs bottom seeded). A smaller size dextran will need to be used in the future to observe differences in permeability. • TEER peaked around day 6 in both static and fluidic conditions, and remained stable throughout the remainder of the culture period (up to day 10). TEER was higher (by ~25 Ohm*cm2) in static cultures compared to fluidic, and was further increased (by 5-10 Ohm*cm2) by seeding cells on the bottom of the membrane (compared to top seeded cells). • Viability (CellTiterGlo) was similar across all seeding conditions. TEX-VAL Courtney Sakolish 4 2023-05-23 TexVal_Consortium 551
View/Edit BIOLOGXsym Exp 3 2021-05-21 TOX LAMPS Fluidic-3D -No Diseases- 301 0 0 0 0 0 0 LAMPS model data generation of Tocilizumab, IL-6, NF and IL-6 + Tocilizumab toxicity modeling UPDDI Larry Vernetti 4 2021-07-06 Taylor_MPS 552
View/Edit BIOLOGXsym Exp 2 2021-05-14 TOX LAMPS Fluidic-3D -No Diseases- 213 0 0 0 0 0 0 LAMPS model for testing GGF2, Tocilizumab and Non Fasting Media Control UPDDI Larry Vernetti 4 2021-07-06 Taylor_MPS 553
View/Edit Lung Study 2 - LPS Exposure (Endothelial) 2020-03-03 TOX Small Airway - ALI (UPenn) Static-3D -No Diseases- 546 0 5 0 0 0 0 This study consists of 12 small airway chips (6 vehicle, 6 LPS treated). The chips were airlifted on day 0, and treatment with LPS (1ug/mL) began on day 11 of airlift. Treatment lasted 7 days, and media was collected and exchanged daily (through the endothelial channel on bottom). Endpoint for this experiment was on day 18. At endpoint, chips were tested for ciliary beating (angle of beating, beat velocity) and Permeability (FITC-Dextran 70kDa). Cell coverage on membranes was also observed via DAPI nuclear staining. TEX-VAL Courtney Sakolish 4 2022-05-13 TexVal_Consortium 554
View/Edit Lung Study 3 - LPS and PM2.5 Exposure (Epithelial) 2020-08-17 TOX Small Airway - ALI (UPenn) Static-3D -No Diseases- 7,971 0 84 0 0 0 0 In this study, 14 chips were seeded with primary small airway cells (SAECs) in the top chamber, and human microvascular endothelial cells (HMVECs) in the bottom chamber. The top channel was airlifted, and the chip was maintained in ALI (air liquid interface) for 15 days prior to treatment. On treatment day, the top chamber was flooded with either untreated media, or media treated with LPS (1ug/mL) or PM2.5 (10ug/mL) for 2 hours. After exposure, media was removed from the top and chips were re-airlifted without rinsing. Chips were allowed to recover 24h until day 16, when endpoint analyses were performed. Endpoints included ciliary beating (velocity), Viability (Calcein AM image quantification), LDH release, and ZO-1 staining quantification (Immunocytochemistry). TEX-VAL Courtney Sakolish 4 2022-05-13 TexVal_Consortium 555
View/Edit BIOLOGXsym Exp 1B 2021-04-26 TOX vLAMPS Fluidic-3D -No Diseases- 33 0 0 0 0 0 0 vLAMPS testing GGF2, Tocilizumab, Non fasting media UPDDI Larry Vernetti 4 2021-07-21 Taylor_MPS 557
View/Edit Lung Study 4 - Open vs Closed Design 2020-10-22 CC Small Airway - ALI (UPenn), Small Airway - ALI (UPenn) - Open Design Fluidic-3D, Static-3D -No Diseases- 286 0 16 0 0 0 0 In this study, 24 chips were seeded with HMVEC (human microvascular endothelial cells) and SAECs (small airway epithelial cells), and brought to airlift. 12 chips were the traditional small airway lung design (OLD), but the other 12 chips had an altered upper layer to allow for direct air exchange over the cells at ALI (NEW). Cultures were taken out over 16 days under static airflow conditions, then moved to an air exposure chamber, where they were exposed to either 0, 5, or 10mL/min clean air-flow for 5 hours, then moved back to normal culturing conditions and allowed to recover overnight until endpoint on day 17. At endpoint, chips were tested for Viability and LDH release, as well as imaging (b-tubulin, ZO-1, cell height, membrane coverage). TEX-VAL Courtney Sakolish 4 2022-05-13 TexVal_Consortium 558
View/Edit Liver_Mimetas 2-lane_Exp.1_iHep and NPCs with Troglitazone 2021-06-27 TOX Liver vs Mimetas 2-lane 2D, Mimetas 2-lane Liver Chip Fluidic-3D, Static-2D -No Diseases- 455 0 606 0 0 0 1 In this study, the hepatotoxicity of the classic hepatotoxic substance troglitazone was evaluated using the Mimetas 2-lane plate model. Before seeding into the Mimetas 2-lane plate, induced pluripotent stem (iPS) human hepatocytes (iHeps) were pre-differentiated in 6-well plates for 5 days. On Day 0, iHeps with collagen were injected into the extracellular matrix (ECM) lanes of Mimetas 2-lane plate. After that, HMEC-1 endothelial cells and THP-1 monocytes were loaded into the perfusion lanes, and DMEM/F12 based media was added to the perfusion lanes. Media was collected and refreshed once every two days. The plate was incubated at 37°C with perfusion by rocking. Cultures were treated with troglitazone (180uM) for 72hrs. After 72hrs troglitazone exposure, cell viability analysis using Propidium Iodide and morphological analysis using immunocytochemistry were conducted. To evaluate hepatotoxicity, albumin, BUN, and LDH in the effluent were measured at 24, 48, and 72 hrs compared to time 0. In addition, the CYP3A4 /Luciferin-IPA assay using cultured cells was conducted before and after 72hrs troglitazone exposure. iHeps with gel were also seeded on a 384-well plate, and troglitazone-induced hepatotoxicity was evaluated under similar conditions. These results were compared with the MPS platform. TEX-VAL Yuki Kato 4 2022-08-31 TexVal_Consortium 566
View/Edit RFE129_NAFLD Drug combination study 2021-07-12 EFF-DM LAMPS Fluidic-3D NAFLD 480 0 0 0 0 0 2 Rationale for drug combination testing: There is currently no single drug for the treatment of NAFLD due in large part to the heterogeneous nature and involvement of multiple pathways in NAFLD disease progression. Our recent computational work has predicted a panel of repurposable drugs that we have begun testing in LAMPS evaluating a panel of metrics. We have tested individual control and predicted drugs and have found that these compounds alleviate differing disease-related phenotypes in our model. The goal now is to test these drugs in combination and determine if multiple disease metrics can be reduced using this combination approach. UPDDI Mark Miedel 4 2023-08-21 Taylor_MPS 578
View/Edit Liver_Mimetas 2-lane_Exp.2_iHep and NPCs with Five Chemical Cocktail 2021-07-08 PK Liver vs Mimetas 2-lane 2D, Mimetas 2-lane Liver Chip Fluidic-3D, Static-2D -No Diseases- 788 0 863 0 0 0 0 In this study, the metabolic capacity with the 5-chemical cocktail was evaluated using the Mimetas 2-lane plate model. Before seeding into the Mimetas 2-lane plate, induced pluripotent stem (iPS) human hepatocytes (iHeps) were pre-differentiated in 6-well plates for 5 days. On Day 0, iHeps with collagen were injected into the extracellular matrix (ECM) lanes of the Mimetas 2-lane plate. After that, HMEC-1 endothelial cells and THP-1 monocytes were loaded into the perfusion lanes, and DMEM/F12 based media was added to the perfusion lanes. Media was collected and refreshed at least once every two days. The plate was incubated at 37°C with perfusion by rocking. Cultures were treated with the 5-chemical cocktail (100uM phenacetin, 20uM coumarin, 10uM diclofenac, 10uM terfenadine, 10uM phenolphthalein) for 24hrs on Days 6, 12, and 16. After 24hrs of exposure, the remaining compound and produced metabolites were measured by LC-MS/MS. Cell viability analysis using Propidium Iodide and morphological analysis using immunocytochemistry were conducted on Day 17. To evaluate basal hepatic function and the hepatotoxicity due to the 5-chemical cocktail, albumin, BUN, and LDH in the effluent were measured on Days 1, 5, 7, 13, and 17. In addition, the CYP3A4 /Luciferin-IPA assay using cultured cells was conducted on Days 6, 7, 16, and 17. iHeps with gel were also seeded on a 384-well plate, and metabolic capacity and basal function were evaluated under similar conditions. These results were compared with the MPS platform. TEX-VAL Yuki Kato 4 2023-05-23 TexVal_Consortium 581
View/Edit Liver_Mimetas 2-lane_Exp.3_iHep and NPCs (Transporters) 2021-07-17 CC Mimetas 2-lane Liver Chip, vs Mimetas 2-lane_2D Fluidic-3D, Static-2D -No Diseases- 587 0 1,119 0 0 0 0 In this study, transporter protein expression/localization and protein production from hepatocytes were evaluated using the Mimetas 2-lane plate model. Before seeding into the Mimetas 2-lane plate, induced pluripotent stem (iPS) human hepatocytes (iHeps) were pre-differentiated in 6-well plates for 5 days. On Day 0, iHeps with collagen were injected into the extracellular matrix (ECM) lanes of the Mimetas 2-lane plate. After that, HMEC-1 endothelial cells and THP-1 monocytes were loaded into the perfusion lanes, and DMEM/F12 based media was added to the perfusion lanes. Media was collected and refreshed once every two days. The plate was incubated at 37°C with perfusion by rocking. No test articles were treated. Cell viability analysis using Propidium Iodide and transporter protein expression using immunocytochemistry were conducted. To evaluate hepatotoxicity, albumin, BUN, Total bilirubin, Total bile acid, AFP, ALT, ALP and LDH in the effluent were measured on 1, 5, 7, 11, and 15. iHeps with gel were also seeded on a 384-well plate and tested under similar conditions. These results were compared with the MPS platform. TEX-VAL Yuki Kato 4 2022-08-31 TexVal_Consortium 582
View/Edit CNBio_Proximal Tubule_Exp3 2021-06-22 CC Proximal Tubule (CNBio - T12), Proximal Tubule (Transwell) Fluidic-3D, Static-3D -No Diseases- 496 0 88 0 0 0 0 In this study, Lonza primary RPTECs were seeded in both the CNBio System (TC12) and Transwell cultures (TW). RPTECs were seeded on either the apical or basolateral surface of the membranes to determine the effects of direct flow on the monocultures. 24 wells/chips were seeded into each growth platform (TC12, TW). 12 wells/chips from each platform were cultured in each seeding configuration (apical or basolateral), and within each of those groups, 6 had media changes every 24h, and the other 6 had media changes every 48h. TC12 cultures were grown under a flow rate of 1uL/s, and TW were static. Cultures were taken out over 17 days, with media changes and TEER measurements throughout. At endpoint, cultures were tested for pAH (para-aminohippuric acid) secretion, permeability testing, and viability testing. Conclusions: • TEER remained low throughout culture period (8-15 Ohm*cm2) with the primary RPTECs compared to previous studies with TERT1-RPRECs, where TEER remained near 100 Ohm*cm2). Maximum TEER was achieved around day 8, and began to drop for the remainder of the culture period until endpoint at day 12. TEER was slightly higher in static cultures, and in bottom seeded vs top seeded cultures, which is concordant with previous studies. • Permeability to FITC-dextran (150kDa) was significantly higher in top seeded cultures compared to bottom seeded, indicating greater barrier function when cells are seeded on the bottom of the membrane. Additionally, static cultures were generally more leaky (higher permeability) than their fluidic counterparts. • Water transport was not observed with these primary cells as previously shown in the TERT lines. In all conditions, volume is increasing in the bottom chamber. Additionally, this observation appears to be cumulative, as the “48hr media change” cultures had greater accumulation of volume in the bottom well than the “24hr media change” groups. • PrestoBlue was used as a non-destructive method for viability testing, and indicated no difference in viability in top vs bottom seeded cultures (which is in agreement with previous studies). Additionally, viability was slightly higher (though not significant) in fluidic cultures compared to static counterparts. • pAH secretion studies showed no difference in cell seeded cultures vs blanks, therefore these cells were not actively secreting pAH, and any transport observed was due to passive diffusion between chambers. Mannitol showed similar trends. • Immunocytochemistry revealed significant differences in monolayer formation between top and bottom seeded cultures. Nuclear staining revealed that when cells were seeded on the top of the membrane, they formed large clusters, with many “holes” in the monolayer. Cells only formed flat monolayers when seeded on the bottom of the membranes. “48h media change” cultures showed more evenly spread monolayers, and better overall density of cells compared to the “24h media change” conditions. Therefore the additional media changes may induce additional stress in cultures. Lastly fluidic cultures showed significantly higher density of cells compared to static conditions. Overall, the best configuration for culturing cells appeared to be bottom seeded, 48h media change conditions in fluidic cultures. • AQP1 expression (confirmed via ICC) was 2-3 fold higher in fluidic conditions compared to static, and media change frequency did not appear to have an effect on AQP1. Top seeded conditions showed higher AQP1 staining, though this did not appear to be significant. • SGLT2 expression (confirmed via ICC) was higher in static conditions compared to fluidic. Media change frequency did not appear to have an effect on SGLT2, however expression was higher in top seeded cultures vs their bottom seeded counterparts. TEX-VAL Courtney Sakolish 4 2023-05-23 TexVal_Consortium 583
View/Edit Liver_Mimetas 2-lane_Exp.4_iHep and NPCs with Five Chemical Cocktail_2nd 2021-08-12 PK Liver vs Mimetas 2-lane 2D, Mimetas 2-lane Liver Chip Fluidic-3D, Static-2D -No Diseases- 928 0 1,816 0 0 0 0 In this study, the metabolic capacity with the 5-chemical cocktail was evaluated using the Mimetas 2-lane plate model. This study is a re-test of Mimetas 2-lane_Exp.2. Before seeding into the Mimetas 2-lane plate, induced pluripotent stem (iPS) human hepatocytes (iHeps) were pre-differentiated in 6-well plates for 5 days. On Day 0, iHeps with collagen were injected into the extracellular matrix (ECM) lanes of the Mimetas 2-lane plate. After that, HMEC-1 endothelial cells and THP-1 monocytes were loaded into the perfusion lanes, and DMEM/F12 based media was added to the perfusion lanes. Media was collected and refreshed at least once every two days. The plate was incubated at 37°C with perfusion by rocking. Cultures were treated with the 5-chemical cocktail (100uM phenacetin, 20uM coumarin, 10uM diclofenac, 10uM terfenadine, 10uM phenolphthalein) for 24hrs on Days 6, 12, and 16. After 24hrs of cocktail exposure, the remaining compound and produced metabolites were measured by LC-MS/MS. Cell viability analysis using Propidium Iodide and morphological analysis using immunocytochemistry were conducted on Day 17. To evaluate basal hepatic function and the hepatotoxicity due to the 5-chemical cocktail, albumin, BUN, and LDH in the effluent were measured on Days 2, 4, 7, 13, and 17. In addition, the CYP3A4 /Luciferin-IPA assay using cultured cells was conducted on Days 4, 7, 13, and 17. iHeps with gel were also seeded on a 384-well plate, and metabolic capacity and basal function were evaluated under similar conditions. These results were compared with the MPS platform. TEX-VAL Yuki Kato 4 2022-08-31 TexVal_Consortium 588
View/Edit Liver_Mimetas 2-lane_Exp.5_PHH (LZ HUM183231) with Midazolam 2021-08-19 PK Mimetas 2-lane Liver Chip Fluidic-3D -No Diseases- 258 0 738 0 0 0 0 In this study, basic hepatic function and metabolic capacity were evaluated to select the optimal hepatocyte batch to use for future MPS platforms. On Day 0, primary human hepatocytes (PHH, Lonza, HUM183231) with collagen were injected into the extracellular matrix (ECM) lanes of the Mimetas 2-lane plate. After that, hepatocyte maintenance media was added to the perfusion lanes. Media was collected and refreshed at least once every two days. The plate was incubated at 37°C with perfusion by rocking. Cultures were treated with midazolam (MDZ, 5uM) for 24hrs on Days 4, 8, and 12. After 24hrs of MDZ exposure, the remaining MDZ and produced metabolites were measured by LC-MS/MS. Cell viability analysis using Propidium Iodide was conducted on Day 14. To evaluate basal hepatic function and the hepatotoxicity due to MDZ, albumin, BUN, and LDH in the effluent were measured on Days 4, 8,12, and 14. In addition, the CYP3A4 /Luciferin-IPA assay using cultured cells was conducted on Days 4, 8, 12, and 14. PHHs were also seeded on a CNBIO LC12 plate and 96-well plate (sandwich culture), and metabolic capacity and basal hepatic function were evaluated under similar conditions. Those results were compared with the Mimetas 2-lane model (see TEX-VAL-PK-2021-08-19-Liver_CNBIO LC12_Exp.3_PHH (LZ HUM183231) with Midazolam). TEX-VAL Yuki Kato 4 2023-05-23 TexVal_Consortium 589
View/Edit Liver_CNBIO LC12_Exp.3_PHH (LZ HUM183231) with Midazolam 2021-08-19 PK CNBIO Liver-MPS (LC12) Consumable Plate, Liver vs CNBIO MPS (LC12) Plate Fluidic-3D, Static-2D -No Diseases- 849 0 226 0 0 0 0 In this study, basic hepatic function and metabolic capacity were evaluated to select the optimal hepatocyte batch to use for future MPS platforms. On Day 0, primary human hepatocytes (PHH, Lonza, HUM183231) were injected into each well of a CNBIO PhysioMimix MPS-LC12 plate with hepatocyte plating media. Media was collected and refreshed at least once every two days (Except on Days 1 to 4 - the same medium was used for 3 days). The plate was incubated at 37°C with the Incubate program (flow rate: 1 µL/s). Cultures were treated with 5 µM midazolam (MDZ) for 24hrs on Days 4, 8, and 12. After 24hrs of MDZ exposure, the remaining MDZ and produced metabolites were measured by LC-MS/MS. Total protein assay was conducted on Day 14 to measure the total protein content of cells in culture. To evaluate basal hepatic function, albumin, BUN, and LDH in the effluent were measured on Days 4, 8,12, and 14. In addition, the CYP3A4/Luciferin-IPA assay using cultured cells was conducted on Days 4, 8, 12, and 14. PHHs were also seeded on a 96-well plate (sandwich culture), and metabolic capacity and basal hepatic function were evaluated under similar conditions. These results were compared with the CNBIO LC12 plate model. (see TEX-VAL-PK-2021-08-19-Liver_Mimetas 2-lane_Exp.5_PHH (LZ HUM183231) with Midazolam). TEX-VAL Yuki Kato 4 2023-05-23 TexVal_Consortium 593
View/Edit Liver_CNBIO LC12_Exp.4_PHH (TF-HU8373) with Midazolam 2021-09-12 PK CNBIO Liver-MPS (LC12) Consumable Plate, Liver vs CNBIO MPS (LC12) Plate Fluidic-3D, Static-2D -No Diseases- 815 0 250 0 0 0 0 In this study, basic hepatic function and metabolic capacity were evaluated to select the optimal hepatocyte batch to use for future MPS platforms. On Day 0, primary human hepatocytes (PHH, Thermo Fisher, HU8373) were injected into each well of a CNBIO PhysioMimix MPS-LC12 plate with William's E medium based media. Media was collected and refreshed at least once every two days (Except on Days 1 to 4 - the same medium was used for 3 days). The plate was incubated at 37°C with the Incubate program (flow rate: 1 µL/s). Cultures were treated with 5 µM midazolam (MDZ) for 24hrs on Days 4, 8, and 12. After 24hrs of MDZ exposure, the remaining MDZ and produced metabolites were measured by LC-MS/MS. Total protein assay was conducted on Day 14 to measure the total protein content of cells in culture. To evaluate basal hepatic function, albumin, BUN, and LDH in the effluent were measured on Days 4, 8,12, and 14. In addition, the CYP3A4/Luciferin-IPA assay using cultured cells was conducted on Days 4, 8, 12, and 14. PHHs were also seeded on a 96-well plate (sandwich culture), and metabolic capacity and basal hepatic function were evaluated under similar conditions. These results were compared with the CNBIO LC12 plate model. TEX-VAL Yuki Kato 4 2023-05-23 TexVal_Consortium 594
View/Edit CNBio_Proximal Tubule_Exp4 (6 cell comparison) 2021-07-20 CC Proximal Tubule (CNBio - T12), Proximal Tubule (Transwell) Fluidic-3D, Static-3D -No Diseases- 2,292 0 612 0 0 0 0 In this study, 6 RPTEC sources were seeded in both the CNBio System (TC12) and Transwell cultures (TW). RPTECs were seeded on either the apical or basolateral surface of the membranes to determine the effects of direct flow on the monocultures. 12 wells/chips were seeded into each growth platform (TC12, TW). 6 wells/chips from each platform were cultured in each seeding configuration (apical or basolateral) - see design image below. TC12 cultures were grown under a flow rate of 2uL/s, and TW were static. Cultures were taken out between 9-18 days (depending on cell source), with media changes and TEER measurements throughout. At endpoint, cultures were tested for pAH (para-aminohippuric acid) secretion, mannitol secretion, permeability testing, and protein expression (Immunocytochemistry, image-based quantification). pAH and mannitol secretion was tested at 1, 2, and 24h after addition to media chambers. Conclusions: • Significant differences in TEER were observed between cell sources. Primary RPTECs (Biopredic and Lonza) remained stable, but low throughout culture period (both ~10 Ω*cm2, up to 14 days in culture). TERT lines were variable – The basal line reached its peak TEER around day 8 (150 Ω*cm2 static, 100 Ω*cm2 fluidic), TERT-OAT1 peaked around day 2-4 (120 Ω*cm2 static and fluidic), TERT-OCT2 peaked around day 8 (30 Ω*cm2 static, 18 Ω*cm2 fluidic), and TERT-OAT3 peaked around day 8 (100 Ω*cm2 static, 80 Ω*cm2 fluidic). Overall, static growth conditions led to higher TEER than fluidic counterparts. Additionally, when cells were seeded on the bottom of the Transwells, TEER was further elevated (by 10-20 Ω*cm2 in TERT parent, OAT1, and OAT3 lines). • Permeability to 70kDa FITC-Dextran was higher in static vs fluidic conditions, and also in top vs bottom seeded conditions. The growth conditions with the tightest barrier to diffusion (lowest dextran permeability) were bottom seeded, fluidic cultures. Within this configuration, all TERT lines showed 1-2% permeability (relative to blanks), and primary cells had 3-4% permeability. To compare, primary cultures in static conditions when grown on the top of the membrane had up to 25% permeability. • Water transport (as observed by differences in ABS) was only observed in the basal TERT1 line, and to a lower degree, in OAT1 lines. Water transport was not observed in the remaining TERT1 lines, or in primary cells. • Immunocytochemistry revealed significant differences in monolayer formation between cell sources. All TERT1 lines showed nice, dense, even monolayers across the membranes in all conditions (top vs bottom; static vs fluidic). However, as previously demonstrated in earlier studies, the primary cells formed large clusters of cell aggregates rather than an even monolayer. • AQP1 expression (confirmed via ICC) was ~30% higher in primary cell cultures compared to TERT1 lines. No significant difference in AQP1 was observed between top/bottom seeding conditions, however AQP1 was markedly higher in fluidic cultures. • SGLT2 expression (confirmed via ICC) was higher in OAT1 cultures, than other cell sources. Additionally, SGLT2 was slightly elevated (though non-significant) in top seeded configurations. No difference was observed in static vs fluidic cultures. • For future studies, RPTECs will be seeded on the bottom of membranes – this appears to result in more even monolayer formation, and higher barrier function over time. Now that a seeding configuration has been determined, future studies can involve compound treatments with more replicates. TEX-VAL Courtney Sakolish 4 2023-05-23 TexVal_Consortium 596
View/Edit Quantification of γH2AX foci per cell 2021-09-22 TOX Salivary Gland [URochester] Static-3D -No Diseases- 15 0 0 0 0 0 0 reatment of Mouse SGm in MB-hydrogels with WR-1065 demonstrates radioprotection from DNA damage after irradiation. Quantification of γH2AX foci per cell SalGlandTC(UR) Elana Amir 4 2022-01-28 Benoit_Rochester 599
View/Edit Quantification of 53BP1 foci per cell 2021-09-21 TOX Salivary Gland [URochester] Static-3D -No Diseases- 15 0 0 0 0 0 0 Treatment of Mouse SGm in MB-hydrogels with WR-1065 demonstrates radioprotection from DNA damage after irradiation. Quantification of 53BP1 foci per cell SalGlandTC(UR) Elana Amir 4 2022-01-28 Benoit_Rochester 601
View/Edit Viability across MB 2021-09-21 TOX Salivary Gland [URochester] Static-3D -No Diseases- 11 0 0 0 0 0 0 Quantification of the percentage of MB containing viable cells at day 2 post- irradiation SalGlandTC(UR) Elana Amir 4 2022-01-28 Benoit_Rochester 602
View/Edit DNA damage after irradiation compared to in vivo response 2021-09-21 TOX Salivary Gland [URochester] Static-3D -No Diseases- 15 0 0 0 0 0 0 DNA damage response after irradiation of SGm in MB-hydrogels compared to in vivo response SalGlandTC(UR) Elana Amir 4 2022-01-28 Benoit_Rochester 603
View/Edit Quantification of MB containing viable cells 2021-09-21 TOX Salivary Gland [URochester] Static-3D -No Diseases- 12 0 0 0 0 0 0 Characterization of salivary gland MB-hydrogel culture system. Quantification of the percentage of MB containing viable cells at days 0, 7, and 14. Statistical analysis was performed using one-way ANOVA with Dunnett’s post hoc test with α = 0.05. N = 3–5, n = 250–290 MB per time point. ns not significant. SalGlandTC(UR) Elana Amir 4 2022-01-28 Benoit_Rochester 606
View/Edit Markers with gene expression of Aqp5 2021-09-21 TOX Salivary Gland [URochester] Static-3D -No Diseases- 16 0 0 0 0 0 0 Mouse SGm in MB-hydrogels express markers of three major cell types in the salivary gland. Gene expression of Aqp5 (a) SalGlandTC(UR) Elana Amir 4 2022-02-11 Benoit_Rochester 612
View/Edit Markers with gene expression of Nkcc1 2021-09-21 TOX Salivary Gland [URochester] Static-3D -No Diseases- 16 0 0 0 0 0 0 Mouse SGm in MB-hydrogels express markers of three major cell types in the salivary gland. Gene expression of Nkcc1 SalGlandTC(UR) Elana Amir 4 2022-02-11 Benoit_Rochester 614
View/Edit Markers with gene expression of Mist1 2021-09-21 TOX Salivary Gland [URochester] Static-3D -No Diseases- 16 0 0 0 0 0 0 Mouse SGm in MB-hydrogels express markers of three major cell types in the salivary gland. Gene expression of Mist1 SalGlandTC(UR) Elana Amir 4 2022-02-11 Benoit_Rochester 615
View/Edit Markers with gene expression of K7 2021-09-21 TOX Salivary Gland [URochester] Static-3D -No Diseases- 16 0 0 0 0 0 0 Mouse SGm in MB-hydrogels express markers of three major cell types in the salivary gland. Gene expression of K7 SalGlandTC(UR) Elana Amir 4 2022-02-11 Benoit_Rochester 616
View/Edit Markers with gene expression of K5 2021-09-21 TOX Salivary Gland [URochester] Static-3D -No Diseases- 16 0 0 0 0 0 0 Mouse SGm in MB-hydrogels express markers of three major cell types in the salivary gland. Gene expression of K5 SalGlandTC(UR) Elana Amir 4 2022-02-11 Benoit_Rochester 617
View/Edit Markers with gene expression of Sma 2021-09-21 TOX Salivary Gland [URochester] Static-3D -No Diseases- 16 0 0 0 0 0 0 Mouse SGm in MB-hydrogels express markers of three major cell types in the salivary gland. Gene expression of Sma SalGlandTC(UR) Elana Amir 4 2022-02-11 Benoit_Rochester 618
View/Edit Gene expression of Pip 2021-09-21 TOX Salivary Gland [URochester] Static-3D -No Diseases- 16 0 0 0 0 0 0 SGm in MB-hydrogels retain expression of secretory proteins. Gene expression of the secretory protein Pip SalGlandTC(UR) Elana Amir 4 2022-02-11 Benoit_Rochester 626
View/Edit Gene expression of amylase 2021-09-21 TOX Salivary Gland [URochester] Static-3D -No Diseases- 7 0 0 0 0 0 0 SGm in MB-hydrogels retain expression of secretory proteins. Gene expression of amylase SalGlandTC(UR) Elana Amir 4 2022-03-01 Benoit_Rochester 627
View/Edit Gene expression of Amy1 2021-09-21 TOX Salivary Gland [URochester] Static-3D -No Diseases- 15 0 0 0 0 0 0 SGm in MB-hydrogels retain expression of secretory proteins. Gene expression of amylase SalGlandTC(UR) Elana Amir 4 2022-01-28 Benoit_Rochester 628
View/Edit Gene expression of Smr3a 2021-09-21 TOX Salivary Gland [URochester] Static-3D -No Diseases- 9 0 0 0 0 0 0 SGm in MB-hydrogels retain expression of secretory proteins. Gene expression of Smr3a SalGlandTC(UR) Elana Amir 4 2022-01-28 Benoit_Rochester 629
View/Edit Gene expression of Ngf 2021-09-21 TOX Salivary Gland [URochester] Static-3D -No Diseases- 12 0 0 0 0 0 0 SGm in MB-hydrogels retain expression of secretory proteins. Gene expression of Ngf SalGlandTC(UR) Elana Amir 4 2022-01-28 Benoit_Rochester 630
View/Edit Gene expression of Muc5b 2021-09-21 TOX Salivary Gland [URochester] Static-3D -No Diseases- 9 0 0 0 0 0 0 SGm in MB-hydrogels retain expression of secretory proteins. Gene expression of Muc5b SalGlandTC(UR) Elana Amir 4 2022-01-28 Benoit_Rochester 631
View/Edit Gene expression of Cst10 2021-09-21 TOX Salivary Gland [URochester] Static-3D -No Diseases- 9 0 0 0 0 0 0 SGm in MB-hydrogels retain expression of secretory proteins. Gene expression of Cst10 SalGlandTC(UR) Elana Amir 4 2022-01-28 Benoit_Rochester 632
View/Edit Gene expression of Cst3 2021-09-21 TOX Salivary Gland [URochester] Static-3D -No Diseases- 9 0 0 0 0 0 0 SGm in MB-hydrogels retain expression of secretory proteins. Gene expression of Cst3 SalGlandTC(UR) Elana Amir 4 2022-01-28 Benoit_Rochester 633
View/Edit Gene expression of Lyz2 2021-09-21 TOX Salivary Gland [URochester] Static-3D -No Diseases- 18 0 0 0 0 0 0 SGm in MB-hydrogels retain expression of secretory proteins. Gene expression of Lyz2 SalGlandTC(UR) Elana Amir 4 2022-01-28 Benoit_Rochester 634
View/Edit Gene expression of M3r 2021-10-25 TOX Salivary Gland [URochester] Static-3D -No Diseases- 16 0 0 0 0 0 0 Mouse SGm in MB-hydrogels are responsive to stimulation with muscarinic agonist carbachol (CCh) and purinergic agonist ATP. Gene expression profiles of muscarinic receptor type 3 (M3r: a) SalGlandTC(UR) Elana Amir 4 2022-02-11 Benoit_Rochester 635
View/Edit Gene expression of P2x7 2021-09-21 TOX Salivary Gland [URochester] Static-3D -No Diseases- 16 0 0 0 0 0 0 Mouse SGm in MB-hydrogels are responsive to stimulation with muscarinic agonist carbachol (CCh) and purinergic agonist ATP. Gene expression profiles of purinergic receptors (P2x7: b) SalGlandTC(UR) Elana Amir 4 2022-02-11 Benoit_Rochester 636
View/Edit Gene expression of P2y2 2021-09-21 TOX Salivary Gland [URochester] Static-3D -No Diseases- 16 0 0 0 0 0 0 Mouse SGm in MB-hydrogels are responsive to stimulation with muscarinic agonist carbachol (CCh) and purinergic agonist ATP. Gene expression profiles of purinergic receptors (P2y2: c) SalGlandTC(UR) Elana Amir 4 2022-02-11 Benoit_Rochester 637
View/Edit Gene expression of acinar marker AQP5 2021-09-21 TOX Salivary Gland [URochester] Static-3D -No Diseases- 12 0 0 0 0 0 0 Human SGm in MB-hydrogels maintain viability and retain markers of functional phenotype. Gene expression of acinar markers AQP5 SalGlandTC(UR) Elana Amir 4 2022-02-14 Benoit_Rochester 639
View/Edit Gene expression of acinar marker NKCC1 2021-09-21 TOX Salivary Gland [URochester] Static-3D -No Diseases- 12 0 0 0 0 0 0 Human SGm in MB-hydrogels maintain viability and retain markers of functional phenotype. Gene expression of acinar marker NKCC1 SalGlandTC(UR) Elana Amir 4 2022-02-14 Benoit_Rochester 640
View/Edit Gene expression of acinar marker MIST1 2021-09-21 TOX Salivary Gland [URochester] Static-3D -No Diseases- 12 0 0 0 0 0 0 Human SGm in MB-hydrogels maintain viability and retain markers of functional phenotype. Gene expression of acinar marker MIST1 SalGlandTC(UR) Elana Amir 4 2022-02-14 Benoit_Rochester 641
View/Edit Fluorescent traces of mouse SGm upon CCh and ATP stimulation 2021-09-15 TOX Salivary Gland [URochester] Static-3D -No Diseases- 5,365 0 0 0 0 0 0 Fig. 4 Mouse SGm in MB-hydrogels are responsive to stimulation with muscarinic agonist carbachol (CCh) and purinergic agonist ATP. Representative fluorescent traces of responsive SGm at day 7 upon CCh and ATP stimulation. NOTE: Due to MPS-Db data formatting requirements, in this study Chips are actually individual microbubbles, and the Groups are the microbubble chips. This ensures that these data are properly aggregated and displayed by MPS-Db. Also, the data presented are comprised of aggregates from all bubble measurements. SalGlandTC(UR) Elana Amir 4 2022-01-28 Benoit_Rochester 653
View/Edit Fluorescent traces of human SGm upon CCh and ATP stimulation at Day 0 2021-09-02 TOX Salivary Gland [URochester] Static-3D -No Diseases- 25,200 0 0 0 0 0 0 Fig. 5 Calcium signaling analysis of AIDUCs from MB-hydrogels at Day 0. CCh and ATP stimulation leads to an elevation of [Ca2+]i in AIDUCs NOTE: Due to MPS-Db data formatting requirements, in this study Chips are actually individual microbubbles, and the Groups are the microbubble chips. This ensures that these data are properly aggregated and displayed by MPS-Db. Also, the data presented are comprised of aggregates from all bubble measurements. SalGlandTC(UR) Elana Amir 4 2022-02-14 Benoit_Rochester 659
View/Edit Fluorescent traces of human SGm upon CCh and ATP stimulation at Day 14 2021-09-06 TOX Salivary Gland [URochester] Static-3D -No Diseases- 28,800 0 0 0 0 0 0 Fig. 5 Calcium signaling analysis of AIDUCs from MB-hydrogels at Day 14. CCh and ATP stimulation leads to an elevation of [Ca2+]i in AIDUCs NOTE: Due to MPS-Db data formatting requirements, in this study Chips are actually individual microbubbles, and the Groups are the microbubble chips. This ensures that these data are properly aggregated and displayed by MPS-Db. Also, the data presented are comprised of aggregates from all bubble measurements. SalGlandTC(UR) Elana Amir 4 2022-02-14 Benoit_Rochester 660
View/Edit Fluorescent traces of human SGm upon CCh and ATP stimulation at Day 7 2021-09-06 TOX Salivary Gland [URochester] Static-3D -No Diseases- 10,200 0 0 0 0 0 0 Fig. 5 Calcium signaling analysis of AIDUCs from MB-hydrogels at Day 7. CCh and ATP stimulation leads to an elevation of [Ca2+]i in AIDUCs NOTE: Due to MPS-Db data formatting requirements, in this study Chips are actually individual microbubbles, and the Groups are the microbubble chips. This ensures that these data are properly aggregated and displayed by MPS-Db. Also, the data presented are comprised of aggregates from all bubble measurements. SalGlandTC(UR) Elana Amir 4 2022-02-14 Benoit_Rochester 661
View/Edit Representative trace of human SGm response to CCh and ATP stimulation at day 7. 2021-09-05 TOX Salivary Gland [URochester] Static-3D -No Diseases- 2,035 0 0 0 0 0 0 Fig. 5 Representative trace of SGm response to CCh (u) and ATP (v) stimulation at day 7. NOTE: Due to MPS-Db data formatting requirements, in this study Chips are actually individual microbubbles, and the Groups are the microbubble chips. This ensures that these data are properly aggregated and displayed by MPS-Db. Also, the data presented are comprised of aggregates from all bubble measurements. SalGlandTC(UR) Elana Amir 4 2022-01-28 Benoit_Rochester 662
View/Edit Mouse SGm in MB-hydrogels with CCh stimulation at Day 0 and changes in [Ca2+]i in AIDUCs 2021-08-26 TOX Salivary Gland [URochester] Static-3D -No Diseases- 3,000 0 0 0 0 0 0 Fig. 4 Mouse SGm in MB-hydrogels are responsive to stimulation with muscarinic agonist carbachol (CCh) and purinergic agonist ATP. CCh stimulation leads to an elevation of [Ca2+]i at day 0 (e) Changes in [Ca2+]i in AIDUCs (h) NOTE: Due to MPS-Db data formatting requirements, in this study Chips are actually individual microbubbles, and the Groups are the microbubble chips. This ensures that these data are properly aggregated and displayed by MPS-Db. Also, the data presented are comprised of aggregates from all bubble measurements. SalGlandTC(UR) Elana Amir 4 2022-02-11 Benoit_Rochester 664
View/Edit CCh stimulation and changes in [Ca2+] in SGm at Day 7 2021-08-04 TOX Salivary Gland [URochester] Static-3D -No Diseases- 138,000 0 0 0 0 0 0 Fig. 4 Mouse SGm in MB-hydrogels are responsive to stimulation with muscarinic agonist carbachol (CCh) and purinergic agonist ATP. CCh stimulation leads to an elevation of [Ca2+] in SGM (day 7: f). Changes in [Ca2+]i in SGm (at day 7: i, NOTE: Due to MPS-Db data formatting requirements, in this study Chips are actually individual microbubbles, and the Groups are the microbubble chips. This ensures that these data are properly aggregated and displayed by MPS-Db. Also, the data presented are comprised of aggregates from all bubble measurements. SalGlandTC(UR) Elana Amir 4 2022-02-11 Benoit_Rochester 665
View/Edit Percent of SGm in MBs responsive to CCh and ATP 2021-09-06 TOX Salivary Gland [URochester] Static-3D -No Diseases- 18 0 0 0 0 0 0 Fig. 4 Mouse SGm in MB-hydrogels are responsive to stimulation with muscarinic agonist carbachol (CCh) and purinergic agonist ATP. m. Percent of SGm in MBs responsive to CCh and ATP SalGlandTC(UR) Elana Amir 4 2022-03-01 Benoit_Rochester 667
View/Edit CNBio_Proximal Tubule_Exp5 (3 Compound Tox/Transport) 2021-12-03 TOX Proximal Tubule (CNBio - T12), Proximal Tubule (Transwell) Fluidic-3D, Static-3D -No Diseases- 5,199 0 474 0 0 0 0 This study consisted of 2 arms – the first arm involved the TERT1/RPTEC-basal line, and the second was with the TERT1/RPTEC-OAT1 line. In each arm of the study, 6 plates were seeded (3 fluidic – CNBio T12; 3 static – Transwell). One plate was for Cisplatin (10uM) exposure, one for Tenofovir (100uM) exposure, and the last plate was for PFOA (1uM) exposure. Cells were cultured in static or fluidic conditions for 10 days, then compound treatment was initiated. Compounds were either added to the top chamber (B->A transport), or the bottom chamber (A->B transport) on days 10, 11, 12, 14, and 16 to determine differences in transport from apical or basolateral sides of the RPTECs. Media was collected on these exposure days to monitor LDH, KIM-1, and transport of compounds across the membrane via analytical chemistry (LCMS for Tenofovir and PFOA; ICP-MS for Cisplatin). TEER was measured throughout culture, and at endpoint, wells were selected for lysis (transciptomics), or permeability (10, 70kDa Dextran) followed by fixation for Immunocytochemistry (ZO-1, OAT1). Conclusions: • Sample analysis currently in progress, however some preliminary data has been analyzed – see below. • TEER peaked for TERT1-basal cells around day 8 (120 Ω*cm2 static, 80 Ω*cm2 fluidic), and day 6 in TERT1-OAT1 cells (160 Ω*cm2 static, 140 Ω*cm2 fluidic). TEER remained constant in the basal line, however began to steadily drop in the OAT1 line over time. As for treatments, in the basal line, TEER was increased significantly when Cisplatin (10uM) was added to the top chamber (basolateral surface of the cells) in fluidic cultures only. No other compound-induced changes in TEER were observed under any culture configuration or compound treatment. These treatments were intended as “non-toxic” so changes in TEER were not expected. • Permeability to 3kDa dextran was higher in all static conditions compared to fluidic (tighter barrier function in fluidic cultures). Compound treatments slightly lowered permeability compared to untreated controls – it is possible the cells were tightening up barriers in response to the treatment with these compounds to reduce passive diffusion. No significant difference was observed from A->B or B->A direction in 3kDa dextran permeability, indicating that this is a passive diffusion based process. Similar patterns were observed in experiments with the 70kDa dextran, however, overall permeability was 2-5 fold lower, indicating that the diffusion rate is size-dependent. • OAT-1 expression (through ICC staining) was higher in fluidic cultures compared to static counterparts in untreated controls. Treatments with the selected compounds/concentrations did not show any clear patterns in changes of OAT-1 expression. • Analytical chemistry, LDH release, KIM-1 secretion, and additional imaging are still in progress, and will provide additional insight. TEX-VAL Courtney Sakolish 4 2023-05-23 TexVal_Consortium 670
View/Edit Liver_CNBIO LC12_Exp.6_PHH (LZ HUM182531) with Midazolam 2021-09-30 PK CNBIO Liver-MPS (LC12) Consumable Plate, Liver vs CNBIO MPS (LC12) Plate Fluidic-3D, Static-2D -No Diseases- 435 0 72 0 0 0 0 In this study, basic hepatic function and metabolic capacity were evaluated to select the optimal hepatocyte batch to use for future MPS platforms. On Day 0, primary human hepatocytes (PHH, Lonza, HUM182531) were injected into each well of a CNBIO PhysioMimix MPS-LC12 plate with hepatocyte plating media. Media was collected and refreshed at least once every two days (Except on Days 1 to 4 - the same medium was used for 3 days). The plate was incubated at 37°C with the Incubate program (flow rate: 1 µL/s). Cultures were treated with 5 µM midazolam (MDZ) for 24hrs on Days 4, 8, and 12. After 24hrs of MDZ exposure, the remaining MDZ and produced metabolites were measured by LC-MS/MS. Total protein assay was conducted on Day 14 to measure the total protein content of cells in culture. To evaluate basal hepatic function, albumin, BUN, and LDH in the effluent were measured on Days 4, 8,12, and 14. In addition, the CYP3A4/Luciferin-IPA assay using cultured cells was conducted on Days 4, 8, 12, and 14. PHHs were also seeded on a 96-well plate (sandwich culture), and metabolic capacity and basal hepatic function were evaluated under similar conditions. These results were compared with the CNBIO LC12 plate model. TEX-VAL Yuki Kato 4 2023-05-23 TexVal_Consortium 678
View/Edit Liver_CNBIO LC12_Exp.5_PHH (TF-HU8300) with Midazolam 2021-09-19 PK CNBIO Liver-MPS (LC12) Consumable Plate, Liver vs CNBIO MPS (LC12) Plate Fluidic-3D, Static-2D -No Diseases- 770 0 130 0 0 0 0 In this study, basic hepatic function and metabolic capacity were evaluated to select the optimal hepatocyte batch to use for future MPS platforms. On Day 0, primary human hepatocytes (PHH, Thermo Fisher, HU8300) were injected into each well of a CNBIO PhysioMimix MPS-LC12 plate with William's E medium based media. Media was collected and refreshed at least once every two days (Except on Days 1 to 4 - the same medium was used for 3 days). The plate was incubated at 37°C with the Incubate program (flow rate: 1 µL/s). Cultures were treated with 5 µM midazolam (MDZ) for 24hrs on Days 4, 8, and 12. After 24hrs of MDZ exposure, the remaining MDZ and produced metabolites were measured by LC-MS/MS. Total protein assay was conducted on Day 14 to measure the total protein content of cells in culture. To evaluate basal hepatic function, albumin, BUN, and LDH in the effluent were measured on Days 4, 8,12, and 14. In addition, the CYP3A4/Luciferin-IPA assay using cultured cells was conducted on Days 4, 8, 12, and 14. PHHs were also seeded on a 96-well plate (sandwich culture), and metabolic capacity and basal hepatic function were evaluated under similar conditions. These results were compared with the CNBIO LC12 plate model. TEX-VAL Yuki Kato 4 2023-05-23 TexVal_Consortium 679
View/Edit Liver_Mimetas 2-lane_Exp.6_iHep ± NPCs with Five Chemical Cocktail 2021-08-30 PK Mimetas 2-lane Liver Chip Fluidic-3D -No Diseases- 300 0 896 0 0 0 0 In this study, the hepatic function and metabolic capacity with the 5-chemical cocktail were evaluated using the Mimetas 2-lane plate model in monoculture of induced pluripotent stem (iPS) human hepatocytes (iHeps) and co-culture of iHeps and non-parenchymal cells (NPCs). Before seeding into the Mimetas 2-lane plate, iHeps were pre-differentiated in 6-well plates for 5 days. On Day 0, iHeps with collagen were injected into the extracellular matrix (ECM) lanes of the Mimetas 2-lane plate. After that, HMEC-1 endothelial cells and THP-1 monocytes were loaded into the perfusion lanes of half the chips in the plate, and DMEM/F12 based media was added to the perfusion lanes of all chips. Media was collected and refreshed at least once every two days. The plate was incubated at 37°C with perfusion by rocking. Cultures were treated with the 5-chemical cocktail (100uM phenacetin, 20uM coumarin, 10uM diclofenac, 10uM terfenadine, 10uM phenolphthalein) for 24hrs on Days 6, 12, and 16. After 24hrs the remaining parent compounds and produced metabolites were measured by LC-MS/MS. Cell viability analysis using Propidium Iodide was conducted on Day 17. To evaluate basal hepatic function, albumin, BUN, and LDH in the effluent were measured on Days 2, 4, 7, 13, and 17. In addition, the CYP3A4/Luciferin-IPA assay using cultured cells was conducted on Days 4, 7, 13, and 17. TEX-VAL Yuki Kato 4 2022-08-31 TexVal_Consortium 680
View/Edit Liver_Mimetas 2-lane_Exp.8_iHep ± NPCs with Midazolam 2021-10-13 PK Mimetas 2-lane Liver Chip Fluidic-3D -No Diseases- 605 0 1,146 0 0 0 0 In this study, the hepatic function and metabolic capacity of both monocultured induced pluripotent stem (iPS) human hepatocytes (iHeps) and iHeps co-cultured with non-parenchymal cells (NPCs) were evaluated in the Mimetas 2-lane plate model with midazolam treatment. Before seeding into the Mimetas 2-lane plate, iHeps were pre-differentiated in 6-well plates for 5 days. On Day 0, iHeps with collagen were injected into the extracellular matrix (ECM) lanes of the Mimetas 2-lane plate. After that, HMEC-1 endothelial cells and THP-1 monocytes were loaded into the perfusion lanes of half the chips in the plate, and DMEM/F12 based media was added to the perfusion lanes of all chips. The media were collected and refreshed at least once every two days. The plate was incubated at 37°C with perfusion by rocking. Cultures were treated with 5 µM midazolam (MDZ) for 24hrs on Days 6, 12, and 16. After 24hrs the remaining MDZ and metabolites produced were measured by LC-MS/MS. Cell viability analysis using Propidium Iodide and morphological analysis using immunocytochemistry were conducted on Day 17. To evaluate basal hepatic function, albumin, BUN, and LDH in the effluent were measured on Days 2, 4, 7, 13, and 17. In addition, the CYP3A4/Luciferin-IPA assay using cultured cells was conducted on Days 7, 13, and 17. TEX-VAL Yuki Kato 4 2022-08-31 TexVal_Consortium 681
View/Edit Liver_Mimetas 2-lane_Exp.9_PHH (TF HU8373) ± NPCs with Midazolam 2021-11-15 PK Liver vs CNBIO MPS (LC12) Plate, Mimetas 2-lane Liver Chip Fluidic-3D, Static-2D -No Diseases- 988 0 1,636 0 0 0 0 In this study, the hepatic function and metabolic capacity of both monocultured primary human hepatocytes (PHHs) and PHHs co-cultured with non-parenchymal cells (NPCs) were evaluated in the Mimetas 2-lane plate model with midazolam treatment. On Day 0, PHHs with collagen were injected into the extracellular matrix (ECM) lanes of the Mimetas 2-lane plate. After that, HMEC-1 endothelial cells and THP-1 monocytes were loaded into the perfusion lanes of half the chips in the plate, and William E based media was added to the perfusion lanes of all chips. The media were collected and refreshed at least once every two days. The plate was incubated at 37°C with perfusion by rocking. Cultures were treated with 5 µM midazolam (MDZ) for 24hrs on Days 6, 12, and 16. After 24hrs the remaining MDZ and metabolites produced were measured by LC-MS/MS. Cell viability analysis using Propidium Iodide and morphological analysis using immunocytochemistry were conducted on Day 17. To evaluate basal hepatic function, albumin, BUN, and LDH in the effluent were measured on Days 2, 4, 7, 13, and 17. In addition, the CYP3A4/Luciferin-IPA assay using cultured cells was conducted on Days 4, 7, 13, and 17. PHHs were also seeded on a 96-well plate (sandwich culture), and metabolic capacity and basal hepatic function were evaluated under similar conditions. Those results were compared with the Mimetas 2-lane model. TEX-VAL Yuki Kato 4 2022-08-31 TexVal_Consortium 696
View/Edit Liver_Mimetas 2-lane_Exp.7_PHH (TF HU8373) pre-differentiated in spheroid plate 2021-11-20 PK Mimetas 2-lane Liver Chip Fluidic-3D -No Diseases- 536 0 1,068 0 0 0 0 In this study, the hepatic function and metabolic capacity of both monocultured primary human hepatocytes (PHHs) and PHHs co-cultured with non-parenchymal cells (NPCs) were evaluated in the Mimetas 2-lane plate model with midazolam treatment. Before seeding into the Mimetas 2-lane plate, PHHs were pre-differentiated in spheroid plates for 7 days. On Day 0, PHHs with collagen were injected into the extracellular matrix (ECM) lanes of the Mimetas 2-lane plate. After that, HMEC-1 endothelial cells and THP-1 monocytes were loaded into the perfusion lanes of half the chips in the plate, and DMEM/F12 based media was added to the perfusion lanes of all chips. The media were collected and refreshed at least once every two days. The plate was incubated at 37°C with perfusion by rocking. Cultures were treated with 5 µM midazolam (MDZ) for 24hrs on Days 6 and 12. After 24hrs the remaining MDZ and metabolites produced were measured by LC-MS/MS. Cell viability analysis using Propidium Iodide and morphological analysis using immunocytochemistry were conducted on Day 14. To evaluate basal hepatic function, albumin, BUN, and LDH in the effluent were measured on Days 2, 4, 7, and 13. In addition, the CYP3A4/Luciferin-IPA assay using cultured cells was conducted on Days 4, 7, and 13. TEX-VAL Yuki Kato 4 2022-08-31 TexVal_Consortium 698
View/Edit Liver_Mimetas 2-lane_Exp.11_iHep and NPCs with Trovafloxacin ± LPS 2021-12-13 TOX Liver vs Mimetas 2-lane 2D, Mimetas 2-lane Liver Chip Fluidic-3D, Static-2D -No Diseases- 538 0 1,080 0 0 0 0 In this study, the immune-mediated hepatotoxicity of the trovafloxacin was evaluated using the Mimetas 2-lane plate model. Before seeding into the Mimetas 2-lane plate, induced pluripotent stem (iPS) human hepatocytes (iHeps) were pre-differentiated in 6-well plates for 5 days. On Day 0, iHeps with collagen were injected into the extracellular matrix (ECM) lanes of Mimetas 2-lane plate. After that, HMEC-1 endothelial cells and THP-1 monocytes were loaded into the perfusion lanes, and DMEM/F12 based media was added to the perfusion lanes. Media was collected and refreshed once every two days. The plate was incubated at 37°C with perfusion by rocking. On Day 8, Cultures were treated with trovafloxacin (25, 100 uM) with and without LPS for 48hrs. After 48hrs trovafloxacin exposure, cell viability analysis using Propidium Iodide and morphological analysis using immunocytochemistry were conducted. To evaluate hepatotoxicity, albumin, BUN, and LDH in the effluent were measured at 48 hrs compared to time 0. In addition, the CYP3A4 /Luciferin-IPA assay using cultured cells was conducted before and after 48hrs trovafloxacin exposure. iHeps with gel were also seeded on a 384-well plate, and trovafloxacin-induced hepatotoxicity was evaluated under similar conditions. These results were compared with the MPS platform. TEX-VAL Yuki Kato 4 2022-08-31 TexVal_Consortium 699
View/Edit Liver_CNBIO LC12_Exp.8_PHH (TF-HU8373) ± THP-1 with LPS 2021-11-24 CC CNBIO Liver-MPS (LC12) Consumable Plate, Liver vs CNBIO MPS (LC12) Plate Fluidic-3D, Static-2D -No Diseases- 286 0 126 0 0 0 0 In this study, we investigated whether primary human hepatocytes (PHHs) and THP-1 monocytes can be co-cultured on CNBIO PhysioMimix MPS-LC12 plate and treated with pro-inflammatory agent (Lipopolysaccharide, LPS) to create an immune-activated liver MPS model. On Day 0, primary human hepatocytes (PHH, Thermo Fisher, HU8373) were cultured into each well of a LC12 plate with William's E medium based media. For the half of the plate, THP1s were also cultured. Media was collected and refreshed at least once every two days (Except on Days 1 to 4). The plate was incubated at 37°C with the Incubate program (flow rate: 1 µL/s). Cultures were treated with 1 ug/mL LPS for 48hrs on Day 8. After 48hrs of LPS exposure, total protein assay was conducted to measure the total protein content of cells in culture. To evaluate basal hepatic function and inflammatory reaction, albumin, BUN, LDH, TNF-alpha, and IL-6 in the effluent were measured on Days 4, 6, 8, and 10. In addition, the CYP3A4/Luciferin-IPA assay using cultured cells was conducted on Days 8 and 10. PHHs with and without THP-1s were also seeded on a 96-well plate (sandwich culture), basal hepatic function and inflammatory reaction were evaluated under similar conditions. These results were compared with the CNBIO LC12 plate model. TEX-VAL Yuki Kato 4 2023-05-23 TexVal_Consortium 701
View/Edit Liver_CNBIO LC12_Exp.9_iHep with Midazolam 2021-11-21 PK CNBIO Liver-MPS (LC12) Consumable Plate, Liver vs CNBIO MPS (LC12) Plate Fluidic-3D, Static-2D -No Diseases- 167 0 132 0 0 0 0 In this study, basic hepatic function and metabolic capacity were evaluated using CNBIO PhysioMimix MPS-LC12 plate with iPSC-derived hepatocytes (iHeps). On Day -5, iHeps were seeded into 6-well plate coated with type 1 collagen with RPMI 1640 medium based media. On Day 0, differentiated iHeps were seeded into each well of a CNBIO PhysioMimix MPS-LC12 plate with DMEM/F12 medium based media. Media was collected and refreshed at least once every two days (Except on Days 1 to 4 - the same medium was used for 3 days). The plate was incubated at 37°C with the Incubate program (flow rate: 1 µL/s). Cultures were treated with 5 µM midazolam (MDZ) for 24hrs on Days 6 and 12. After 24hrs of MDZ exposure, the remaining MDZ and produced metabolites were measured by LC-MS/MS. Total protein assay was conducted on Day 14 to measure the total protein content of cells in culture. To evaluate basal hepatic function, albumin, BUN, and LDH in the effluent were measured on Days 4, 7,12, and 14. In addition, the CYP3A4/Luciferin-IPA assay using cultured cells was conducted on Days 4, 7, and 13. iHeps were also seeded on a 96-well plate (sandwich culture), and metabolic capacity and basal hepatic function were evaluated under similar conditions. These results were compared with the CNBIO LC12 plate model. TEX-VAL Yuki Kato 4 2023-05-23 TexVal_Consortium 702
View/Edit Gene expression of muscarinic receptor M3R 2022-01-20 TOX Salivary Gland [URochester] Static-3D -No Diseases- 12 0 0 0 0 0 0 Human SGm in MB-hydrogels maintain viability and retain markers of functional phenotype. Gene expression of muscarinic receptor M3R SalGlandTC(UR) Elana Amir 4 2022-02-14 Benoit_Rochester 703
View/Edit Gene expression of purinergic receptor P2X7 2022-01-13 TOX Salivary Gland [URochester] Static-3D -No Diseases- 12 0 0 0 0 0 0 Human SGm in MB-hydrogels maintain viability and retain markers of functional phenotype. Gene expression of purinergic receptor P2X7 SalGlandTC(UR) Elana Amir 4 2022-02-14 Benoit_Rochester 704
View/Edit Gene expression of purinergic receptor P2Y2 2022-01-13 TOX Salivary Gland [URochester] Static-3D -No Diseases- 12 0 0 0 0 0 0 Human SGm in MB-hydrogels maintain viability and retain markers of functional phenotype. Gene expression of purinergic receptor P2Y2 SalGlandTC(UR) Elana Amir 4 2022-02-14 Benoit_Rochester 705
View/Edit Quantification of Human SGm in MB-hydrogels viability 2022-01-13 TOX Salivary Gland [URochester] Static-3D -No Diseases- 9 0 0 0 0 0 0 Fig. 5 Human SGm in MB-hydrogels maintain viability and retain markers of functional phenotype. LIVE/DEAD assay of human chips quantification (d). Scale bar = 500 μm. Statistical analysis was performed using one-way ANOVA with Dunnett’s post hoc test. SalGlandTC(UR) Elana Amir 4 2022-03-01 Benoit_Rochester 706
View/Edit Quantification of percent responsive microbubbles per chip. 2022-01-13 TOX Salivary Gland [URochester] Static-3D -No Diseases- 15 0 0 0 0 0 0 Fig. 5 Human SGm in MB-hydrogels maintain viability and retain markers of functional phenotype. u–w Whole chip calcium assay via fluorescently labeled calcium-binding indicator Calbryte 520 AM. u and v Representative trace of SGm response to CCh (u) and ATP (v) stimulation at day 7. w Quantification of percent responsive microbubbles per chip. Data are represented as fluorescent intensity (f) divided by fluorescence at time 0 (fo). Percent of SGm in MBs responsive to CCh and ATP (w). SalGlandTC(UR) Elana Amir 4 2022-03-01 Benoit_Rochester 707
View/Edit CCh stimulation and changes in [Ca2+] in SGm at Day 14 2022-01-01 TOX Salivary Gland [URochester] Static-3D -No Diseases- 57,600 0 0 0 0 0 0 Fig. 4 Mouse SGm in MB-hydrogels are responsive to stimulation with muscarinic agonist carbachol (CCh) and purinergic agonist ATP. CCh stimulation leads to an elevation of [Ca2+]I at day 14: g) Changes in [Ca2+]i in AIDUCs and in SGm day 14: j NOTE: Due to MPS-Db data formatting requirements, in this study Chips are actually individual microbubbles, and the Groups are the microbubble chips. This ensures that these data are properly aggregated and displayed by MPS-Db. Also, the data presented are comprised of aggregates from all bubble measurements. SalGlandTC(UR) Elana Amir 4 2022-02-11 Benoit_Rochester 726
View/Edit Liver_Mimetas 2-lane_Exp.12_iHep ± NPCs with Midazolam_2nd 2021-11-30 PK Mimetas 2-lane Liver Chip, vs Mimetas 2-lane_2D Fluidic-3D, Static-2D -No Diseases- 681 0 1,068 0 0 0 0 In this study, the hepatic function and metabolic capacity of both monocultured induced pluripotent stem (iPS) human hepatocytes (iHeps) and iHeps co-cultured with non-parenchymal cells (NPCs) were evaluated in the Mimetas 2-lane plate model with midazolam (MDZ) treatment. This study is a re-test of “Liver_Mimetas 2-lane_Exp.8_iHep ± NPCs with Midazolam”. Before seeding into the Mimetas 2-lane plate, iHeps were pre-differentiated in 6-well plates for 5 days. On Day 0, iHeps with collagen were injected into the extracellular matrix (ECM) lanes of the Mimetas 2-lane plate. After that, HMEC-1 endothelial cells and THP-1 monocytes were loaded into the perfusion lanes of half the chips in the plate, and DMEM/F12 based media was added to the perfusion lanes of all chips. The media were collected and refreshed at least once every two days. The plate was incubated at 37°C with perfusion by rocking. Cultures were treated with 5 µM MDZ for 24hrs on Days 6, 12, and 16. After 24hrs the remaining MDZ and metabolites produced were measured by LC-MS/MS. Cell viability analysis using Propidium Iodide and morphological analysis using immunocytochemistry were conducted on Day 17. To evaluate basal hepatic function, albumin, BUN, and LDH in the effluent were measured on Days 2, 4, 7, 13, and 17. In addition, the CYP3A4/Luciferin-IPA assay using cultured cells was conducted on Days 4, 7, 13, and 17. iHeps with gel were also seeded on a 384-well plate, and metabolic capacity and basal function were evaluated under similar conditions. These results were compared with the MPS platform. TEX-VAL Yuki Kato 4 2022-08-31 TexVal_Consortium 727
View/Edit Liver_CNBIO LC12_Exp.10 PHH (TF-HU8373) with Midazolam_2nd 2021-11-29 PK CNBIO Liver-MPS (LC12) Consumable Plate, Liver vs CNBIO MPS (LC12) Plate Fluidic-3D, Static-2D -No Diseases- 655 0 214 0 0 0 0 In this study, basic hepatic function and metabolic capacity were evaluated to evaluate the reproducibility of previous study. On Day 0, primary human hepatocytes (PHH, Thermo Fisher, HU8373) were injected into each well of a CNBIO PhysioMimix MPS-LC12 plate with William's E medium based media. Media was collected and refreshed at least once every two days (Except on Days 1 to 4 - the same medium was used for 3 days). The plate was incubated at 37°C with the Incubate program (flow rate: 1 µL/s). Cultures were treated with 5 µM midazolam (MDZ) for 24hrs on Days 6, 12, and 16. After 24hrs of MDZ exposure, the remaining MDZ and produced metabolites were measured by LC-MS/MS. Total protein assay was conducted on Day 17 to measure the total protein content of cells in culture. To evaluate basal hepatic function, albumin, BUN, and LDH in the effluent were measured on Days 4, 7,13, and 17. In addition, the CYP3A4/Luciferin-IPA assay using cultured cells was conducted on Days 4, 7, 13, and 17. PHHs were also seeded on a 96-well plate (sandwich culture), and metabolic capacity and basal hepatic function were evaluated under similar conditions. These results were compared with the CNBIO LC12 plate model. TEX-VAL Yuki Kato 4 2023-05-23 TexVal_Consortium 728
View/Edit Kidney Study 13 - Glomerulus - NovaBiosis cells 2021-04-02 CC Glomerulus (Mimetas 3-lane) Fluidic-3D -No Diseases- 214 0 40 0 0 0 0 In this study, glomerular cells were seeded into the Mimetas 3-lane platform in 2 different configurations. The cells used in this study were human primary-cryopreserved Glomerular Endothelial Cells (Novabiosis-NYD-GEC), primary-cryopreserved Podocytes (Celprogen-1314184), and primary-cryopreserved Mesangial Cells (Novabiosis-#001). In configuration 1, all cell types were seeded into the top channel, and the bottom channel was left blank. In configuration 2, podocytes and mesangial cells were seeded into the top channel, and glomerular endothelial cells into the bottom channel. 4mg/mL collagen I gel was used as the barrier in the central channel of all chips. Cells were cultured under rocking conditions (7°, 8 min intervals) for 4 days before endpoint. Phase contrast images were collected daily throughout culture, and on the final day of culture, permeability was assessed by adding FITC-Dextran (150kDa) or FITC-hBSA to the top channel and monitoring fluorescence in the effluent of the bottom channel after 1 hr. Conclusions: • Cells seemed to grow with high viability in both seeding configurations, however a good barrier to dextran and BSA was not established. • Further optimization is necessary for these cells to establish barrier function and glomerular basal membrane (GBM) secretion. • Timing is crucial in the seeding of these cells – too short of a growth period does not allow for barrier formation, however cells quickly begin to invade into the matrix, making longer-term experiment a challenge. TEX-VAL Courtney Sakolish 4 2023-05-23 TexVal_Consortium 733
View/Edit Kidney Study 14 - Glomerulus - ScienCell Cells 2021-04-09 CC Glomerulus (Mimetas 3-lane) Fluidic-3D -No Diseases- 20 0 130 0 0 0 0 In this study, glomerular cells were seeded into the Mimetas 3-lane platform in 2 different configurations. The cells used in this study were human primary-cryopreserved Glomerular Endothelial Cells (ScienCell Research Laboratories-31529), primary-cryopreserved Podocytes (Celprogen-1314184), and primary-cryopreserved Mesangial Cells (ScienCell Research Laboratories-11008). In configuration 1, all cell types were seeded into the top channel, and the bottom channel was left blank. In configuration 2, podocytes and mesangial cells were seeded into the top channel, and glomerular endothelial cells into the bottom channel. 4mg/mL collagen I gel was used as the barrier in the central channel of all chips. Cells were cultured under rocking conditions (7°, 8 min intervals) for 3 days before endpoint. Phase contrast images were collected daily throughout culture, and on the final day of culture, permeability was assessed by adding FITC-Dextran (150kDa) or FITC-hBSA to the top channel and monitoring fluorescence in the effluent of the bottom channel after 1 hr. Additionally, viability testing was performed (image based: Nuclear staining, PI) to determine effects of seeding configuration on cell viability. Conclusions: • Cells seemed to grow with high viability in both seeding configurations, however a good barrier to dextran and BSA was not established. • Further optimization is necessary for these cells to establish barrier function and glomerular basal membrane (GBM) secretion. • Viability was similar between seeding configurations. • Permeability was lower (better barrier function) in configuration 2, where the cells were seeded in the 2 separate channels. However, permeability was still near blank (cell-free) levels, so cells were not providing a good barrier under either condition. • Timing is crucial in the seeding of these cells – too short of a growth period does not allow for barrier formation, however cells quickly begin to invade into the matrix, making longer-term experiment a challenge. TEX-VAL Courtney Sakolish 4 2023-05-23 TexVal_Consortium 734
View/Edit Kidney Study 15 - Glomerulus - ScienCell Cells +/- Mesangial Cells 2021-04-26 CC Glomerulus (Mimetas 3-lane) Fluidic-3D -No Diseases- 19 0 60 0 0 0 0 In this study, glomerular cells were seeded into the Mimetas 3-lane platform in 2 different seeding configurations, and with or without mesangial cells (4 total combinations). The cells used in this study were human primary-cryopreserved Glomerular Endothelial Cells (ScienCell Research Laboratories-31529), primary-cryopreserved Podocytes (Celprogen-1314184), and primary-cryopreserved Mesangial Cells (ScienCell Research Laboratories-11008). In configuration 1, all cell types were seeded into the top channel, and the bottom channel was left blank. In configuration 2, podocytes and mesangial cells were seeded into the top channel, and glomerular endothelial cells into the bottom channel. Within each of these 2 configurations, podocytes and glomerular endothelial cells were either seeded with, or without mesangial cells. 4mg/mL collagen I gel was used as the barrier in the central channel of all chips. Cells were cultured under rocking conditions (7°, 8 min intervals) for only 1 day before endpoint due to high invasion into the gel. Phase contrast images were collected, and on the final day of culture, permeability was assessed by adding FITC-Dextran (150kDa) to the top channel and monitoring fluorescence in the effluent of the bottom channel after 1 hr (plate reader based). Additionally, cells were fixed and immunocytochemistry was performed to determine the presence and location of glomerular (CD34, Nephrin) and endothelial markers (CD31), and glomerular basal membrane secretion (laminin, Collagen IVa1,2, Collagen IVa4). Unfortunately high background fluorescence due to the presence of collagen in the gel layer made interpretation of these images challenging. Conclusions: • Cells seemed to grow with high viability in all seeding configurations, however a good barrier to dextran and BSA was not established. • Further optimization is necessary for these cells to establish barrier function and glomerular basal membrane (GBM) secretion. • Cells invaded into the gel within 24 hours of seeding, making endpoint measurement difficult. • Timing is crucial in the seeding of these cells – too short of a growth period does not allow for barrier formation, however cells quickly begin to invade into the matrix, making longer-term experiment a challenge. • Collagen in the gel lane leads to high background when imaging for other basement membrane markers. TEX-VAL Courtney Sakolish 4 2023-05-23 TexVal_Consortium 735
View/Edit Kidney Study 16 - Glomerulus - NovaBiosis cells, new lot 2021-05-06 CC Glomerulus (Mimetas 3-lane) Fluidic-3D -No Diseases- 20 0 100 0 0 0 0 In this study, glomerular cells were seeded into the Mimetas 3-lane platform in 2 different seeding configurations, and with or without mesangial cells (4 total combinations). The cells used in this study were human primary-cryopreserved Glomerular Endothelial Cells (Novabiosis-NYD-GEC), primary-cryopreserved Podocytes (Novabiosis-FZY-P), and primary-cryopreserved Mesangial Cells (Novabiosis-#001). In configuration 1, all cell types were seeded into the top channel, and the bottom channel was left blank. In configuration 2, podocytes and mesangial cells were seeded into the top channel, and glomerular endothelial cells into the bottom channel. Within each of these 2 configurations, podocytes and glomerular endothelial cells were either seeded with, or without mesangial cells. 4mg/mL collagen I gel was used as the barrier in the central channel of all chips. Cells were cultured under rocking conditions (7°, 8 min intervals) for 3 days before endpoint, however significant gel invasion was observed by the end of this experiment. Phase contrast images were collected on days 2 and 3, and on the final day of culture, permeability was assessed by adding FITC-Dextran (150kDa) to the top channel and monitoring fluorescence in the effluent of the bottom channel after 1 hr (plate reader based). Additionally, viability testing was performed (image based: Nuclear staining, PI) to determine effects of seeding configuration on cell viability. Conclusions: • Cells seemed to grow with high viability in all seeding configurations, however a good barrier to dextran and BSA was not established. • Further optimization is necessary for these cells to establish barrier function and glomerular basal membrane (GBM) secretion. • Timing is crucial in the seeding of these cells – too short of a growth period does not allow for barrier formation, however cells quickly begin to invade into the matrix, making longer-term experiments challenging. TEX-VAL Courtney Sakolish 4 2023-05-23 TexVal_Consortium 737
View/Edit Kidney Study 17 - RPTEC/HUVEC 3-lane Mimetas Analytical Study (Lonza donor 1) 2021-09-30 CC Proximal Tubule/Vascular (Mimetas 3-lane) Fluidic-3D -No Diseases- 970 0 490 0 0 0 0 In this study, 2 duplicate Mimetas 3-lane plates seeded with Lonza RPTECs (lot #18TL117405, top channel) and HUVECs (bottom channel) were exposed to cisplatin (10uM), tenofovir (100uM), PFOA (1uM), or were left untreated. Compounds were added either through the RPTEC or HUVEC channel to measure permeability in both directions, and determine if the RPTECs would preferentially uptake/secrete the compounds. Additionally, some chips were left “cell free” to act as blanks to compare against passive diffusion through the gel. Cells were cultured over 6 days before initiation of treatment. Media was sampled after 1, 2, 3, 5, and 7 days of treatment (total of 13 days culture), and compounds were refreshed with each media change. In addition to analytical chemistry (to monitor permeability of the compounds through the gel), media was collected and tested for LDH and KIM-1. Lastly, viability testing (image based) was performed at endpoint to determine effects of compound treatments on the RPTECs and HUVECs. One plate was treated with lysis buffer for later analysis into transcriptomics, and one plate was fixed for imaging, though due to high background fluorescence in the gel lane, and significant cell invasion, results were inconclusive. Conclusions: • This lot of RPTECs (#18TL117405) showed significant invasion into the gel by day 6 when compound treatment was initiated. This experiment will be repeated with another lot of Lonza primary cells, as invasion into the gel will certainly affect permeability. • Analytical data confirmed that the gel provided a significant barrier to permeability of the tested compounds. Even after being exposed to treatments for 1 week (day 7 exposure), ~30% cisplatin, ~12% PFOA, and ~8% tenofovir was able to pass through the gel of blank, cell-free chips. The addition of RPTECs and HUVECs to the cell channels decreased permeability by 2-10% on average, but it was not possible to determine uptake vs secretion in this system, as equilibrium was never reached through the gel layer in the middle. • LDH testing in effluent showed that when added to the RPTEC channel, cisplatin caused large increases in LDH release in RPTECs, and a slight increase in HUVECs on the opposite side of the gel. Other treatments (Tenofovir, PFOA) did not affect LDH release in either channel at the tested concentrations. When added to the HUVEC channel, cisplatin caused a similar decrease in LDH release (likely due to acute cell death) in both RPTEC and HUVEC channels. Other treatments did not cause changes in LDH release compared to controls. • KIM-1 was increased slightly in all treatment conditions (Cisplatin, Tenofovir, PFOA; added through either RPTEC or HUVEC channel) compared to controls, except for when cisplatin was added directly to the RPTEC channel. In this treatment condition, KIM-1 release decreased significantly after 1 day of exposure, and continued to drop throughout additional exposure days, indicating a decrease in viability of the RPTECs, which were no longer present to secrete this marker. • Viability testing (image based; Nuclear stain + PI) indicated that when Cisplatin was added to the RPTEC channel, viability decreased in both channels (65% viability in RPTEC channel; 16% in HUVEC channel compared to untreated controls). When added to the HUVEC channel, a significant decrease in viability was only observed in the HUVEC channel (20% viability), indicating that HUVECs were sensitive to cisplatin treatment, regardless of where in the chip it was added. Tenofovir decreased viability in RPTECs by ~10% (when added to either channel), and decreased viability in HUVECs by ~15% (when added to either channel). Lastly, PFOA did not significantly affect viability in any condition. • The gel layer continues to be an issue for permeability, making transport studies challenging in this platform. TEX-VAL Courtney Sakolish 4 2023-05-23 TexVal_Consortium 738
View/Edit Kidney Study 18 - RPTEC/HUVEC 3-lane Mimetas Analytical Study (Lonza donor 2) 2021-10-12 CC Proximal Tubule/Vascular (Mimetas 3-lane) Fluidic-3D -No Diseases- 851 0 531 0 0 0 0 This is a re-run of “Kidney Study 17”, using a second donor of primary RPTECs. In this study, 2 duplicate Mimetas 3-lane plates seeded with Lonza RPTECs (lot #18TL114340, top channel) and HUVECs (bottom channel) were exposed to cisplatin (10uM), tenofovir (100uM), PFOA (1uM), or were left untreated. Compounds were added either through the RPTEC or HUVEC channel to measure permeability in both directions, and determine if the RPTECs would preferentially uptake/secrete the compounds. Additionally, some chips were left “cell free” to act as blanks to compare against passive diffusion through the gel. Cells were cultured over 6 days before initiation of treatment. Media was sampled after 1, 2, 3, 5, and 7 days of treatment (total of 13 days culture), and compounds were refreshed with each media change. In addition to analytical chemistry (to monitor permeability of the compounds through the gel), media was collected and tested for LDH and KIM-1. Note that cisplatin analytical samples were not tested due to high operational cost of ICP-MS/MS, and the lack of usable data in the PFOA and Tenofovir samples. Lastly, viability testing (image based) was performed at endpoint to determine effects of compound treatments on the RPTECs and HUVECs. One plate was treated with lysis buffer for later analysis into transcriptomics, and one plate was fixed for imaging, though due to high background fluorescence in the gel lane, results were inconclusive. Conclusions: • This lot of RPTECs (#18TL114340) did not invade into the gel like the previous lot tested, even by endpoint at day 13. • Analytical data confirmed that the gel provided a significant barrier to permeability of the tested compounds. Even after being exposed to treatments for 1 week (day 7 exposure), ~7% PFOA, and ~20% tenofovir was able to pass through the gel of blank, cell-free chips (Cisplatin analysis is still in progress). The addition of RPTECs and HUVECs to the cell channels decreased permeability by 2-10% on average, but it was not possible to determine uptake vs secretion in this system, as equilibrium was never reached through the gel layer in the middle. • LDH testing in effluent showed that when added to the RPTEC channel, cisplatin caused large increases in LDH release in RPTECs, but did not affect HUVECs on the opposite side of the gel. Other treatments (Tenofovir, PFOA) did not affect LDH release in either channel at the tested concentrations. Other treatments did not cause changes in LDH release compared to controls. • KIM-1 was increased slightly in all treatment conditions (Cisplatin, Tenofovir, PFOA; added through either RPTEC or HUVEC channel) compared to controls, except for when cisplatin was added directly to the RPTEC channel. In this treatment condition, KIM-1 release decreased significantly after 1 day of exposure, and continued to drop throughout additional exposure days, indicating a decrease in viability of the RPTECs, which were no longer present to secrete this marker (these results match the previous study with donor #18TL117405). • Viability testing (image based; Nuclear stain + PI) indicated that when Cisplatin was added to the RPTEC channel, viability decreased in both channels (65% viability in RPTEC channel; 14% in HUVEC channel compared to untreated controls). When added to the HUVEC channel, a significant decrease in viability was only observed in the HUVEC channel (17% viability), indicating that HUVECs were sensitive to cisplatin treatment, regardless of where in the chip it was added. These results are nearly identical to the previous study (kidney study 17) with the alternative donor. However, in this study, tenofovir did not affect viability of the RPTECs (when added to either channel), but had a greater effect on the HUVECs (69% viability when added through RPTEC channel; 46% when added directly through HUVEC channel), though standard deviation was high on these measurements. Lastly, PFOA did not significantly affect viability in RPTECs or HUVECs under any condition. • The gel layer continues to be an issue for permeability, making transport studies challenging in this platform. TEX-VAL Courtney Sakolish 4 2023-05-23 TexVal_Consortium 739
View/Edit Gene Expression Array Studies 2021-11-08 CC-OMICS Skeletal Muscle MPS E-stim, Skeletal Muscle MPS non-stim Fluidic-3D -No Diseases- 0 990 0 0 0 0 1 Malany (UF) Zon Thwin 1 Malany_skeletal muscle MPS 750
View/Edit Muscle activation in G-node-integrated tissues_Firing Spontaneous activities at G-node 2020-10-20 CC Muscle activation in geometrically insulated cardiac tissues node Static-2D -No Diseases- 31 0 0 0 0 0 0 Muscle activation in G-node-integrated tissues. Design (top), Ca2+ propagation signals (middle), and activation map (bottom). G-node integration in rectangular tissue with sharp and rounded corners. G-nodes integrated into a rectangular tissue predominantly activated the muscle construct compared to the four corners of the muscle tissue. Furthermore, the rounded corner decreased the empirical probability of initial activation at the corner, and thus the activation probability of the integrated G-node in the rounded tissue increased to 91%. Different shapes of G-node in rounded rectangular tissue. The square and diamond G-nodes have a similar probability of activation at the G-node to the circular design (83 and 87% vs. 91%). This indicates that an acute angles in the small source tissue like the G-node is not critical in determining the activation site. Rather, it is the fewer number of cells in the smaller G-node tissue that acts as a source to initiate muscle activation. DBG Gilyong Lee 4 2022-06-06 Parker_DBG 755
View/Edit Liver_Mimetas 2-lane_Exp.13_iHep and NPCs with Trovafloxacin ± LPS_2nd 2022-02-11 TOX Liver vs Mimetas 2-lane 2D, Mimetas 2-lane Liver Chip Fluidic-3D, Static-2D -No Diseases- 482 0 796 0 0 0 0 In this study, the immune-mediated hepatotoxicity of the trovafloxacin was evaluated using the Mimetas 2-lane plate model. This study is a re-test of Mimetas 2-lane_Exp.11. Before seeding into the Mimetas 2-lane plate, induced pluripotent stem (iPS) human hepatocytes (iHeps) were pre-differentiated in 6-well plates for 5 days. On Day 0, iHeps with collagen were injected into the extracellular matrix (ECM) lanes of Mimetas 2-lane plate. After that, HMEC-1 endothelial cells and THP-1 monocytes were loaded into the perfusion lanes, and DMEM/F12 based media was added to the perfusion lanes. Media was collected and refreshed once every two days. The plate was incubated at 37°C with perfusion by rocking. On Day 8, Cultures were treated with trovafloxacin (50, 150 uM) with and without LPS for 48hrs. After 48hrs trovafloxacin exposure, cell viability analysis using Propidium Iodide and morphological analysis using immunocytochemistry were conducted. To evaluate hepatotoxicity, albumin, BUN, and LDH in the effluent were measured at 48 hrs compared to time 0. In addition, the CYP3A4 /Luciferin-IPA assay using cultured cells was conducted before and after 48hrs trovafloxacin exposure. iHeps with gel were also seeded on a 384-well plate, and trovafloxacin-induced hepatotoxicity was evaluated under similar conditions. These results were compared with the MPS platform. TEX-VAL Yuki Kato 4 2022-08-31 TexVal_Consortium 759
View/Edit Gene expression of acinar cell marker Mist1 2022-03-03 TOX Salivary Gland [URochester] Static-3D -No Diseases- 8 0 0 0 0 0 0 Figure 4. Hydrogel encapsulated SGm express markers of all three major cell types in the salivary gland (acinar, duct, and myoepithelial). Quantitative PCR was used to measure gene expression of acinar cell markers A) Mist 1 SalGlandTC(UR) Elana Amir 4 2022-04-29 Benoit_Rochester 762
View/Edit Gene expression of acinar cell marker Nkcc1 2022-03-03 TOX Salivary Gland [URochester] Static-3D -No Diseases- 8 0 0 0 0 0 0 Gene expression of acinar cell marker Nkcc1. Figure 4. Hydrogel encapsulated SGm express markers of all three major cell types in the salivary gland (acinar, duct, and myoepithelial). Quantitative PCR was used to measure gene expression of acinar cell markers B) Nkcc1 SalGlandTC(UR) Elana Amir 4 2022-04-29 Benoit_Rochester 763
View/Edit Gene expression of acinar cell marker Aqp5 2022-03-03 TOX Salivary Gland [URochester] Static-3D -No Diseases- 8 0 0 0 0 0 0 Gene expression of acinar cell marker Aqp5 Figure 4. Hydrogel encapsulated SGm express markers of all three major cell types in the salivary gland (acinar, duct, and myoepithelial). Quantitative PCR was used to measure gene expression of acinar cell markers C) Aqp5 SalGlandTC(UR) Elana Amir 4 2022-04-29 Benoit_Rochester 764
View/Edit Gene expression of acinar cell marker Ip3r3 2022-03-03 TOX Salivary Gland [URochester] Static-3D -No Diseases- 14 0 0 0 0 0 0 Gene expression of acinar cell marker Ip3r3 Figure 4. Hydrogel encapsulated SGm express markers of all three major cell types in the salivary gland (acinar, duct, and myoepithelial). Quantitative PCR was used to measure gene expression of acinar cell markers D) Ip3r3 SalGlandTC(UR) Elana Amir 4 2022-04-29 Benoit_Rochester 765
View/Edit Gene expression of duct cell marker K7 2022-03-03 TOX Salivary Gland [URochester] Static-3D -No Diseases- 16 0 0 0 0 0 0 Gene expression of duct cell marker K7 Figure 4. Hydrogel encapsulated SGm express markers of all three major cell types in the salivary gland (acinar, duct, and myoepithelial). Quantitative PCR was used to measure gene expression of E) duct cell markers K7 SalGlandTC(UR) Elana Amir 4 2022-04-29 Benoit_Rochester 766
View/Edit Gene expression of duct cell marker K5 2022-03-03 TOX Salivary Gland [URochester] Static-3D -No Diseases- 6 0 0 0 0 0 0 Gene expression of duct cell marker K5 Figure 4. Hydrogel encapsulated SGm express markers of all three major cell types in the salivary gland (acinar, duct, and myoepithelial). Quantitative PCR was used to measure gene expression of duct cell marker F) K5 SalGlandTC(UR) Elana Amir 4 2022-04-29 Benoit_Rochester 767
View/Edit Gene expression of myoepithelial cell marker Sma 2022-03-03 TOX Salivary Gland [URochester] Static-3D -No Diseases- 8 0 0 0 0 0 0 Gene expression of myoepithelial cell marker Sma Figure 4. Hydrogel encapsulated SGm express markers of all three major cell types in the salivary gland (acinar, duct, and myoepithelial). Quantitative PCR was used to measure gene expression of myoepithelial cell marker G) Sma relative to the reporter gene Rps29 (ribosomal gene). SalGlandTC(UR) Elana Amir 4 2022-04-29 Benoit_Rochester 768
View/Edit Gene expression of secretory protein Amy1 2022-03-03 TOX Salivary Gland [URochester] Static-3D -No Diseases- 8 0 0 0 0 0 0 Figure 6. Hydrogel encapsulated SGm retain expression of secretory proteins. Gene expression of the secretory proteins A)Amy1 determined using qPCR SalGlandTC(UR) Elana Amir 4 2022-04-29 Benoit_Rochester 769
View/Edit Gene expression of secretory protein Pip 2022-03-03 TOX Salivary Gland [URochester] Static-3D -No Diseases- 8 0 0 0 0 0 0 Gene expression of secretory protein Pip. Figure 6. Hydrogel encapsulated SGm retain expression of secretory proteins. Gene expression of the secretory proteins B)Pip determined using qPCR SalGlandTC(UR) Elana Amir 4 2022-04-29 Benoit_Rochester 770
View/Edit Gene expression of secretory protein Muc5b 2022-03-03 TOX Salivary Gland [URochester] Static-3D -No Diseases- 16 0 0 0 0 0 0 Gene expression of secretory protein Muc5b Figure 6. Hydrogel encapsulated SGm retain expression of secretory proteins. Gene expression of the secretory proteins C)Muc5b determined using qPCR SalGlandTC(UR) Elana Amir 4 2022-04-29 Benoit_Rochester 771
View/Edit Gene expression of secretory protein Lyz2 2022-03-03 TOX Salivary Gland [URochester] Static-3D -No Diseases- 16 0 0 0 0 0 0 Gene expression of secretory protein Lyz2 Figure 6. Hydrogel encapsulated SGm retain expression of secretory proteins. Gene expression of the secretory proteins D) Lyz2 determined using qPCR SalGlandTC(UR) Elana Amir 4 2022-04-29 Benoit_Rochester 772
View/Edit Gene expression profiles of muscarinic receptor type 3 M3r 2022-03-03 TOX Salivary Gland [URochester] Static-3D -No Diseases- 8 0 0 0 0 0 0 Figure7. SGm in hydrogels are responsive to stimulation with muscarinic agonist carbachol (CCh) and purinergic agonist ATP. Gene expression profiles of muscarinic receptor type 3 (M3r, A) in SGm at days 7 and 14 SalGlandTC(UR) Elana Amir 4 2022-04-29 Benoit_Rochester 774
View/Edit Gene expression profile of purinergic receptor P2y2 2022-03-03 TOX Salivary Gland [URochester] Static-3D -No Diseases- 8 0 0 0 0 0 0 Figure7. SGm in hydrogels are responsive to stimulation with muscarinic agonist carbachol (CCh) and purinergic agonist ATP. Gene expression profiles of purinergic receptor (P2y2, B) in SGm at days 7 and 14 SalGlandTC(UR) Elana Amir 4 2022-04-29 Benoit_Rochester 775
View/Edit Muscle activation in G-node-integrated tissues_Firing Spontaneous activities at top-left 2020-10-20 CC Muscle activation in geometrically insulated cardiac tissues node Static-2D -No Diseases- 31 0 0 0 0 0 0 Muscle activation in G-node-integrated tissues. Design (top), Ca2+ propagation signals (middle), and activation map (bottom). G-node integration in rectangular tissue with sharp and rounded corners. G-nodes integrated into a rectangular tissue predominantly activated the muscle construct compared to the four corners of the muscle tissue. Furthermore, the rounded corner decreased the empirical probability of initial activation at the corner, and thus the activation probability of the integrated G-node in the rounded tissue increased to 91%. Different shapes of G-node in rounded rectangular tissue. The square and diamond G-nodes have a similar probability of activation at the G-node to the circular design (83 and 87% vs. 91%). This indicates that an acute angles in the small source tissue like the G-node is not critical in determining the activation site. Rather, it is the fewer number of cells in the smaller G-node tissue that acts as a source to initiate muscle activation. DBG Gilyong Lee 4 2022-06-06 Parker_DBG 783
View/Edit Muscle activation in G-node-integrated tissues_Firing Spontaneous activities at bottom-left 2020-10-20 CC Muscle activation in geometrically insulated cardiac tissues node Static-2D -No Diseases- 31 0 0 0 0 0 0 Muscle activation in G-node-integrated tissues. Design (top), Ca2+ propagation signals (middle), and activation map (bottom). G-node integration in rectangular tissue with sharp and rounded corners. G-nodes integrated into a rectangular tissue predominantly activated the muscle construct compared to the four corners of the muscle tissue. Furthermore, the rounded corner decreased the empirical probability of initial activation at the corner, and thus the activation probability of the integrated G-node in the rounded tissue increased to 91%. Different shapes of G-node in rounded rectangular tissue. The square and diamond G-nodes have a similar probability of activation at the G-node to the circular design (83 and 87% vs. 91%). This indicates that an acute angles in the small source tissue like the G-node is not critical in determining the activation site. Rather, it is the fewer number of cells in the smaller G-node tissue that acts as a source to initiate muscle activation. DBG Gilyong Lee 4 2022-06-06 Parker_DBG 784
View/Edit Muscle activation in G-node-integrated tissues_Firing Spontaneous activities at top-right 2020-10-20 CC Muscle activation in geometrically insulated cardiac tissues node Static-2D -No Diseases- 31 0 0 0 0 0 0 Muscle activation in G-node-integrated tissues. Design (top), Ca2+ propagation signals (middle), and activation map (bottom). G-node integration in rectangular tissue with sharp and rounded corners. G-nodes integrated into a rectangular tissue predominantly activated the muscle construct compared to the four corners of the muscle tissue. Furthermore, the rounded corner decreased the empirical probability of initial activation at the corner, and thus the activation probability of the integrated G-node in the rounded tissue increased to 91%. Different shapes of G-node in rounded rectangular tissue. The square and diamond G-nodes have a similar probability of activation at the G-node to the circular design (83 and 87% vs. 91%). This indicates that an acute angles in the small source tissue like the G-node is not critical in determining the activation site. Rather, it is the fewer number of cells in the smaller G-node tissue that acts as a source to initiate muscle activation. DBG Gilyong Lee 4 2022-06-06 Parker_DBG 785
View/Edit Muscle activation in G-node-integrated tissues_Firing Spontaneous activities at bottom-right 2020-10-20 CC Muscle activation in geometrically insulated cardiac tissues node Static-2D -No Diseases- 31 0 0 0 0 0 0 Muscle activation in G-node-integrated tissues. Design (top), Ca2+ propagation signals (middle), and activation map (bottom). G-node integration in rectangular tissue with sharp and rounded corners. G-nodes integrated into a rectangular tissue predominantly activated the muscle construct compared to the four corners of the muscle tissue. Furthermore, the rounded corner decreased the empirical probability of initial activation at the corner, and thus the activation probability of the integrated G-node in the rounded tissue increased to 91%. Different shapes of G-node in rounded rectangular tissue. The square and diamond G-nodes have a similar probability of activation at the G-node to the circular design (83 and 87% vs. 91%). This indicates that an acute angles in the small source tissue like the G-node is not critical in determining the activation site. Rather, it is the fewer number of cells in the smaller G-node tissue that acts as a source to initiate muscle activation. DBG Gilyong Lee 4 2022-06-06 Parker_DBG 786
View/Edit Contractility Study of Young Athletic and Old Sedentary Skeletal Muscle Cells 2021-11-08 CC Skeletal Muscle MPS E-stim, Skeletal Muscle MPS non-stim Fluidic-3D -No Diseases- 2,525 0 0 0 0 0 0 Malany (UF) Zon Thwin 1 Malany_skeletal muscle MPS 787
View/Edit Quantification of sphere size 2022-03-17 TOX Salivary Gland [URochester] Static-3D -No Diseases- 49 0 0 0 0 0 0 Figure 2. Cell viability of hydrogel encapsulated AIDUCs. G) Quantification of sphere size at different time points after encapsulation. SalGlandTC(UR) Elana Amir 4 2022-04-29 Benoit_Rochester 789
View/Edit Viability of hydrogel encapsulated AIDUCs 2022-03-17 TOX Salivary Gland [URochester] Static-3D -No Diseases- 18 0 0 0 0 0 0 Figure 2. Cell viability of hydrogel encapsulated AIDUCs. H) Viability of hydrogel encapsulated AIDUCs. ** p < 0.01, **** p < 0.0001 compared with day 0 determined using one-way ANOVA with Dunnett’s correction for multiple comparisons. N = 4. SalGlandTC(UR) Elana Amir 4 2022-04-29 Benoit_Rochester 790
View/Edit Quantitative analysis of Ca2+ signaling in SGm stimulated with CCh 2022-03-17 TOX Salivary Gland [URochester] Static-3D -No Diseases- 539 0 0 0 0 0 0 Figure 7. Quantitative analysis of Ca2+ signaling in SGm stimulated with H) CCh and L) ATP. Dashed lines indicate that the stimuli were added at 60 s and removed at 240 s. All gene expression data is relative to day 0 with ribosomal gene Rps29. Statistics are relative to Day 0, using one-way ANOVA with Dunnett’s post-hoc test. **** p < 0.0001. N = 4. SalGlandTC(UR) Elana Amir 4 2022-04-29 Benoit_Rochester 791
View/Edit Quantitative analysis of Ca2+ signaling in SGm stimulated with ATP 2022-03-18 TOX Salivary Gland [URochester] Static-3D -No Diseases- 585 0 0 0 0 0 0 Cloned from Study SalGlandTC(UR)-TOX-2022-03-17-Quantitative analysis of Ca2+ signaling in SGm stimulated with CCh. Figure 7. Quantitative analysis of Ca2+ signaling in SGm stimulated with H) CCh and L) ATP. Dashed lines indicate that the stimuli were added at 60 s and removed at 240 s. All gene expression data is relative to day 0 with ribosomal gene Rps29. Statistics are relative to Day 0, using one-way ANOVA with Dunnett’s post-hoc test. **** p < 0.0001. N = 4. SalGlandTC(UR) Elana Amir 4 2022-04-29 Benoit_Rochester 795
View/Edit Intraventricular pressure and volume measured by catheterization of Cor.4U based ventricles. Isoproterenol-induced stroke work reduction. 2016-02-02 EFF-CC Tissue Engineered Ventricle Static-3D -No Diseases- 9 0 0 0 0 0 0 Isoproterenol dose-response applied to tissue-engineered ventricles seeded with 3 million Cor.4U stem cell derived cardiomyocytes (approximate cell density of 100,000 per square centimeter of ventricle surface) per ventricle. Intra-ventricular pressure-volume measurements were performed at resting beat rate (0 isoproterenol) and for subsequent increasing doses of isoproterenol. Beat rate and ventricular work were both calculated as a function of isoproterenol dose. Beat rates were acquired from either intra-ventricular pressure or volume measurements. Work performed by ventricle contraction was calculated from the pressure-volume loop area. Intraventricular pressure and volume measured by catheterization of Cor.4U based ventricles. Exposure to isoproterenol (Iso) reduced stroke work. Here, pressure and volume were normalized by polynomial fit to remove measurement drift occurring over the course of multiple Iso doses.*P< 0.05; P values were P = 0.038 (Cor.4U, N= 3 ventricles). Our observation of isoproterenol-induced stroke work reduction is consistent with negative forceñfrequency relationships observed in tissues based on immature cardiomyocyte DBG Albert Cordoves 4 2022-06-06 Parker_DBG 797
View/Edit Isoproterenol-dependent beat rates for human Cor.4U tissue engineered ventricles 2016-02-02 EFF-CC Tissue Engineered Ventricle Static-3D -No Diseases- 32 0 0 0 0 0 0 Isoproterenol dose-response applied to tissue-engineered ventricles seeded with 3 million Cor.4U stem cell derived cardiomyocytes (approximate cell density of 100,000 per square centimeter of ventricle surface) per ventricle. Intra-ventricular pressure-volume measurements were performed at resting beat rate (0 isoproterenol) and for subsequent increasing doses of isoproterenol. Beat rate and ventricular work were both calculated as a function of isoproterenol dose. Beat rates were acquired from either intra-ventricular pressure or volume measurements. Work performed by ventricle contraction was calculated from the pressure-volume loop area. Isoproterenol-dependent beat rates for human Cor.4U based (N= 4) ventricles. Time-domain recordings of chamber volume were Fourier-transformed (FFT) to obtain beat rates. The spontaneous beat rate of Cor.4U ventricles (~85 ± 15 bpm)increased by ~40% following exposure to 10-4 M Iso. *P< 0.05, **P< 0.001, compared with baseline (no Iso), one-way ANOVA with Tukey post-hoc test. Exact P values for Iso-dependent beat rates (Hz) were P= 0.524 (Cor.4U, N= 4 ventricles). For Cor.4U, differences in beat rate were not statistically significant (P= 0.183). In all cases, measurements were performed on day 14 of culture in the tissue engineered ventricle. DBG Albert Cordoves 4 2022-06-06 Parker_DBG 835
View/Edit Inhibition of mTOR signaling enhances maturation of cardiomyocytes derived from human-induced pluripotent stem cells via p53-induced quiescence 2019-08-01 DM 2D on 96 Well Plate Static-2D -No Diseases- 4,368 0 0 0 0 0 0 Tested whether the mTOR inhibitor, Torin1, can promote maturation of stem cell-derived cardiomyocytes. DBG Albert Cordoves 4 2022-06-06 Parker_DBG 837
View/Edit Kidney Study 19 - Mimetas new gel formulation 2021-11-23 CC Mimetas 3-lane Organoplate Fluidic-3D -No Diseases- 34 0 6 0 0 0 0 In this study, we attempted a new gel formulation based on previously published work in another tissue chip model (PMID: 30833403). One of the main issues we have been facing with the Mimetas 3-lane plate is the lack of permeability of compounds/drugs to the opposite channel. Instead of using the 4mg/mL collagen type 1 recommended by Mimetas, we attempted a gel made from 1% gelatin, 25mg/mL fibrinogen, 2.5mM CaCl2 and 0.2% transglutaminase in hopes that this would increase permeability. We were able to successfully polymerize the new gel, however the new formulation did not improve permeability compared to the original formulation. Additionally, the longer incubation time of the new formulation caused some of the original gel lanes to dry up prior to testing permeability. Here, we show the results with phase contrast imaging, the addition of trypan blue to track leakage across the gel, and permeability of FITC-dextrans (10, 70, 150kDa) and FITC-BSA. TEX-VAL Courtney Sakolish 4 2023-05-23 TexVal_Consortium 864
View/Edit Kidney Study 20 - Mimetas agarose gel ECM 2021-11-30 CC Mimetas 3-lane Organoplate Fluidic-3D -No Diseases- 0 0 5 0 0 0 0 In this study, we attempted a few new gel formulations based on fibrinogen or agarose as suggested by the Consortium group. One of the main issues we have been facing with the Mimetas 3-lane plate is the lack of permeability of compounds/drugs to the opposite channel. Instead of using the 4mg/mL collagen type 1 recommended by Mimetas, we attempted to load chips with gels made from 0.2% or 0.5% agarose, or 5 or 25mg/mL fibrinogen in hopes that this would increase permeability. We were able to successfully polymerize the new gels, however the new formulations did not improve permeability compared to the original formulation, and success rate was low. Only 5 chips (1 from each gel loading condition are shown here -- see images). Note: This study has images only, no quantitative data was collected due to low success rate. Conclusions: -The 0.2 and 0.5% agarose gels had negligible differences in gel permeability from the original formulation, however they were significantly more difficult to load into chips, as they had to be kept near-boiling, and would typically stiffen in the pipette before even being loaded into the gel lanes. -The 5mg/mL fibrinogen gel was not stiff enough to hold up in the ECM lane, and collapsed when buffer was added. -The 25mg/mL fibrinogen gel was stiff enough to create an ECM barrier, but permeability was lower than the original collagen gel formulation. TEX-VAL Courtney Sakolish 4 2023-05-23 TexVal_Consortium 865
View/Edit Kidney Study 21 - Mimetas Efflux Transporter Activity - TERT-OAT1 2022-01-21 CC Kidney Proximal Tubule [384-well Static], Proximal Tubule/Vascular (Mimetas 3-lane) Fluidic-3D, Static-2D -No Diseases- 72 0 238 0 0 0 0 In this study, a Mimetas 3-lane plate was seeded in one of 2 configurations. RPTEC only – TERT1-OAT1 RPTECs in the top channel, and bottom channel left empty; or RPTEC + HUVEC – TERT1-OAT1 RPTECs in the top channel, and HUVECs in the bottom channel. After cells reached confluency in both channels (3 days of rocking culture at 7 degrees, 8 minute intervals), chips were treated with CalceinAM, CMFDA, or 6-NBDG in the presence or absence of inhibitors (CyclosporineA, MK571, or Phlorizin, respectively) to measure influx/efflux transporter activity. After incubation with fluorophores +/- inhibitors, RPTEC channels were imaged to determine intracellular fluorescence levels. A 384 well plate was seeded with RPTECs only and used as a 2D comparator. TEX-VAL Courtney Sakolish 4 2023-05-23 TexVal_Consortium 866
View/Edit Kidney Study 22 - Mimetas Efflux Transporter Activity - TERT-parent 2022-02-18 CC Kidney Proximal Tubule [384-well Static], Proximal Tubule/Vascular (Mimetas 3-lane) Fluidic-3D, Static-2D -No Diseases- 96 0 200 0 0 0 0 In this study, a Mimetas 3-lane plate was seeded in one of 2 configurations. RPTEC only – TERT1-Parent RPTECs in the top channel, and bottom channel left empty; or RPTEC + HUVEC – TERT1-parent RPTECs in the top channel, and HUVECs in the bottom channel. After cells reached confluency in both channels (3 days of rocking culture at 7 degrees, 8 minute intervals), chips were treated with CalceinAM, CMFDA, or 6-NBDG in the presence or absence of inhibitors (CyclosporineA, MK571, or Phlorizin, respectively) to measure influx/efflux transporter activity. After incubation with fluorophores +/- inhibitors, RPTEC channels were imaged to determine intracellular fluorescence levels. A 384 well plate was seeded with RPTECs only and used as a 2D comparator. TEX-VAL Courtney Sakolish 4 2023-05-23 TexVal_Consortium 869
View/Edit Kidney Study 23 - Mimetas Efflux Transporter Activity - TERT-OCT2 2022-02-21 CC Kidney Proximal Tubule [384-well Static], Proximal Tubule/Vascular (Mimetas 3-lane) Fluidic-3D, Static-2D -No Diseases- 96 0 200 0 0 0 0 In this study, a Mimetas 3-lane plate was seeded in one of 2 configurations. RPTEC only – TERT1-OCT2 RPTECs in the top channel, and bottom channel left empty; or RPTEC + HUVEC – TERT1-OCT2 RPTECs in the top channel, and HUVECs in the bottom channel. After cells reached confluency in both channels (3 days of rocking culture at 7 degrees, 8 minute intervals), chips were treated with CalceinAM, CMFDA, or 6-NBDG in the presence or absence of inhibitors (CyclosporineA, MK571, or Phlorizin, respectively) to measure influx/efflux transporter activity. After incubation with fluorophores +/- inhibitors, RPTEC channels were imaged to determine intracellular fluorescence levels. A 384 well plate was seeded with RPTECs only and used as a 2D comparator. TEX-VAL Courtney Sakolish 4 2023-05-23 TexVal_Consortium 873
View/Edit Kidney Study 24 - Mimetas Efflux Transporter Activity - Lonza 1 - 18TL117405 2022-03-11 CC Kidney Proximal Tubule [384-well Static], Proximal Tubule/Vascular (Mimetas 3-lane) Fluidic-3D, Static-2D -No Diseases- 59 0 122 0 0 0 0 In this study, a Mimetas 3-lane plate was seeded with Primary RPTECs from Lonza in the top channel and HUVECs in the opposite channel. Half of the plate was seeded with RPTECs from Lot # 18TL117405 (This study), and the other half of the plate was seeded at a later date with RPTECs from lot # 18TL114340 (study 24a). After cells reached confluency in both channels (4 days of rocking culture at 7 degrees, 8 minute intervals), chips were treated with CalceinAM, CMFDA, or 6-NBDG in the presence or absence of inhibitors (CyclosporineA, MK571, or Phlorizin, respectively) to measure influx/efflux transporter activity. After incubation with fluorophores +/- inhibitors, RPTEC channels were imaged to determine intracellular fluorescence levels. A 384 well plate was seeded with the same lot of RPTECs and used as a 2D comparator. TEX-VAL Courtney Sakolish 4 2023-05-23 TexVal_Consortium 874
View/Edit Kidney Study 24a - Mimetas Efflux Transporter Activity - Lonza 2 - 18TL114340 2022-03-15 CC Proximal Tubule/Vascular (Mimetas 3-lane) Fluidic-3D -No Diseases- 17 0 40 0 0 0 0 In this study, a Mimetas 3-lane plate was seeded with Primary RPTECs from Lonza in the top channel and HUVECs in the opposite channel. Half of the plate was seeded with RPTECs from Lot # 18TL114340 (This study), and the other half of the plate was previously seeded at an earlier date with RPTECs from lot # 18TL117405 (study 24). After cells reached confluency in both channels (6 days of rocking culture at 7 degrees, 8 minute intervals), chips were treated with Calcein AM, CMFDA, or 6-NBDG in the presence or absence of inhibitors (Cyclosporine A, MK571, or Phlorizin, respectively) to measure influx/efflux transporter activity. After incubation with fluorophores +/- inhibitors, RPTEC channels were imaged to determine intracellular fluorescence levels. **NOTE: No 384 well plate comparator was seeded in this study due to the slow growth and low cell count on the date of seeding.** TEX-VAL Courtney Sakolish 4 2023-05-23 TexVal_Consortium 876
View/Edit Kidney Study 25 - Mimetas Efflux Transporter Activity - TERT-OAT3 2022-05-02 CC Kidney Proximal Tubule [384-well Static], Proximal Tubule/Vascular (Mimetas 3-lane) Fluidic-3D, Static-2D -No Diseases- 51 0 118 0 0 0 0 In this study, a Mimetas 3-lane plate was seeded in one of 2 configurations. RPTEC only – TERT1-OAT3 RPTECs in the top channel, and bottom channel left empty; or RPTEC + HUVEC – TERT1-OAT3 RPTECs in the top channel, and HUVECs in the bottom channel. After cells reached confluency in both channels (3 days of rocking culture at 7 degrees, 8 minute intervals), chips were treated with CalceinAM, CMFDA, or 6-NBDG in the presence or absence of inhibitors (CyclosporineA, MK571, or Phlorizin, respectively) to measure influx/efflux transporter activity. After incubation with fluorophores +/- inhibitors, RPTEC channels were imaged to determine intracellular fluorescence levels. A 384 well plate was seeded with RPTECs only and used as a 2D comparator. TEX-VAL Courtney Sakolish 4 2023-05-23 TexVal_Consortium 885
View/Edit Tier1_2D_and_3D_Bone-TempO-Seq 2017-08-08 TOX-EFF-DM-CC-OMICS Bone, Bone [TAMU Static] Static-2D, Static-3D Ewing's Sarcoma 0 251,790 0 0 0 0 0 Cloned from Study TEX-VAL-TOX-DM-CC-2017-08-08-Tier1_3D_Bone, then added experimental models from Tier1_2D to add the TempO-Seq results. In this study, 96 scaffolds were shipped to TAMU from the developer lab. 48 of the bone scaffolds contained only human osteoblasts (TE-Bone), and 48 of the scaffolds contained both human osteoblasts as well as Ewing's sarcoma cell aggregates (TE-Tumor). These TE-Bone and TE-Tumor scaffolds were treated with 4.16uM Doxorubicin, 12uM Linsitinib, or their associated controls (0.04% or 0.12% DMSO, respectively). 3D scaffolds were treated in a realistic regimen over either 2 weeks (Doxorubicin) or 3 weeks (Linsitinib). Media was collected daily and tested for osteopontin (OPN) and LDH. At endpoint, scaffolds were either lysed for viability testing (total luminescence), or fixed and sectioned for immunocytochemistry (H&E). TEX-VAL Sandra Karcher 4 2022-09-07 Rusyn_TAMU 914
View/Edit testing constructs 2023-01-04 EFF Demo Plate Model Static-2D -No Diseases- 0 0 0 0 0 0 1 my test study CTDS Andrea Tentner 1 CTDS_HEAL 981
View/Edit Characterization of catecholaminergic polymorphic ventricular tachycardia (CPVT) opto-muscular thin films (MTFs) 2017-07-01 DM Human Engineered CPVT Myocardium Static-3D Catecholaminergic Polymorphic Ventricular Tachycardia (CPVT) 976 0 0 0 0 0 0 DBG Albert Cordoves 4 2023-04-07 Parker_DBG 1045
View/Edit Ca2+ calmodulin–dependent protein kinase II (CaMKII) inhibition suppresses the catecholaminergic polymorphic ventricular tachycardia (CPVT) arrhythmic phenotype 2017-07-01 DM Human Engineered CPVT Myocardium Static-3D Catecholaminergic Polymorphic Ventricular Tachycardia (CPVT) 1,199 0 0 0 0 0 0 Occurrence of reentry in CPVT1 and CPVT2 engineered tissues treated with a CaMKII inhibitor (AIP) DBG Albert Cordoves 4 2023-04-07 Parker_DBG 1047