Fig. 6

Induction of the genes for cytochrome P450 in organoid-derived epithelial cells (A) Expression of the genes for cytochrome P450 (CYP1A1, CYP2B6, CYP2D6, CYP3A4, CYP2C9, and CYP2C19). The expression levels of adult small intestine whole tissue were set to 1.0. Expression levels was calculated from the results of independent (biological) triplicate experiments. Results are expressed as mean ± SD (n = 3). ES-LONG: human ESC-derived mesenchymal stromal cells, ES-RYU: human ESC-derived intestinal epithelial cells, iPS-LONG: human iPSC-derived mesenchymal stromal cells, iPS-RYU: human iPSC-derived intestinal epithelial cells. (B) Induction of the cytochrome P450 genes with exposure to Omeprazole (OM), β-Naphthoflavone (BNF), Phenobarbital (PB), Rifampicin (Rif), vitamin D (VD3), and Dexamethasone (Dex). Results are expressed as mean ± SD (n = 3). The expression level of each gene without any treatment (DMSO) was set to 1.0. Expression levels were calculated from the results of independent (biological) triplicate experiments. Statistical significance was determined using Student's t-test; ** Fold > 2, p < 0.01, * Fold > 2, p < 0.05. (C) Immunocytochemistry of RYU with antibodies to CYP3A4 and CES2, which are drug-metabolizing enzymes. Nuclei were stained with DAPI. Scale bars: 50 µm. (D) Measurement of CYP3A4 activity. The levels of 6β-hydroxy testosterone were measured. Results are expressed as mean ± SD (n = 3 triplicate biological experiments). Statistical significance was determined using Dunnett's test