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. 2020 Apr 6;10:5989. doi: 10.1038/s41598-020-63151-z

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Development of long-term culture method of hiPSCs-IECs. (A) Schematic procedure for long-term culture of hiPSCs-IECs. (B) Bright field images of hiPSC-IECs on days 0, 19 and 54 are shown. Scale bars represent 500 µm. (C) Enterocyte marker VILLIN gene expression in intestinal organoids and long-term cultured hiPSC-IECs are at similar levels. Data are presented as means ± S.D. (intestinal organoids: n = 19, long-term cultured hiPSC-IECs: n = 3) (D) Drug metabolic activity for terfenadine in 8-weeks-cultured iPSC-IECs. Data are presented as means ± S.D. (n = 3). (E) Drug metabolic activity for midazolam in 8-weeks-cultured iPSC-IECs. Data are presented as means ± S.D. (n = 3). ^*p < 0.01 versus the reaction with 10 µM ketoconazole. (F) Tight-junction maker (OCLN and ZO-1) gene expressions in hiPSC-IECs at day 32 post-seeding onto Transwell inserts after 8-weeks-cultured (n = 3). (G) Representative immuno-staining images of OCCLUDIN and ZO-1 in 8-weeks-cultured hiPSC-IEC monolayer after seeding on a culture plate. A scale bar represents 200 µm.