Enteroid cysts (three-dimensional [3D] structures) are seeded onto permeable membrane supports and grown to confluency to generate monolayers (two-dimensional [2D] structures). The resulting monolayers allow for controllable access to both apical and basolateral surfaces of the intestinal epithelial cells.The monolayer format, as opposed to the classical Matrigel-embedded 3D cyst culture, allows manipulations both at the apical and basolateral cell surfaces in a compartmentalized manner, thereby broadening options for treatment and collection of specimens (i.e. cells and culture media). It also improves reproducibility in evaluation of outcomes by reducing the variation in cell number and the restricted lumen volume inherent in the cysts cultures.
This model represents the RED assay conditions and protocol used when testing serum compound binding for the media used in JHU's Intestinal Enteroid model.
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