Development and characterization of a mouse epithelial cell monolayer platform (APECCS) of the small and large intestines

We describe the development and characterization of a mouse epithelial cell monolayer platform of the small and large intestines with a broad range of potential applications including the discovery and development of minimally systemic drug candidates. Culture conditions for each intestinal segment were optimized by correlating monolayer global gene expression with the corresponding tissue segment. The monolayers polarized, formed tight junctions, and contained a diversity of intestinal epithelial cell lineages. Overall design: We established monolayer cultures were with the growth factor combination used for three-dimensional mouse colon organoids i.e. Wnt3a, epidermal growth factor (EGF), noggin, and R-spondin 1 (WENR). RNA sequencing was used to evaluate whether the cultures maintained segment-specific gene expression patterns by comparing the expression of genes with at least five-fold differential expression between mouse proximal and distal colon tissues to the relative expression of these genes in the monolayer cultures (Table S1).