Fig. 2

Optimal medium for monolayer culture of ELC-org (ELC-org-mono). (A) The gene expression levels of drug transporters (MDR1, BCRP, PEPT1), drug-metabolizing enzymes (CYP3A4, UGT1A1, CES1, CES2), intestinal cell markers (CDX2, VIL, MUC2, LYZ, LGR5, EPCAM) and a mesenchymal cell marker (VIM) were examined in ELC-org and ELC-org-mono cultured with different media and in the human small intestine by qRT-PCR. The organoid culture medium (OCM) was the same as the medium previously used for the organoid culture, while intestinal maturation medium (IMM) was the same as the final medium previously used for the differentiation of ELCs differentiation from human iPS cells. Details are shown in the Materials and Methods section. The GAPDH expression level was taken as 1.0. (B) The TEER values in each cell monolayer cultured on cell culture inserts were measured by Millicell-ERS2. (C) The CYP3A4 activity in each cell monolayer was examined by using the P450-Glo CYP3A4 assay kit in the presence or absence of 10 µM ketoconazole (a CYP3A4 inhibitor). All data represent the mean ± S.D. (n = 3, biological replicates). Statistical analyses were performed using the unpaired two-tailed Student’s t-test (***p < 0.005). N.S. means “Not Significant”