Generation of intestinal organoids derived from human pluripotent stem cells for drug testing

被引：58

作者：

Yoshida, Shinpei ^{[1
,3
]}

Miwa, Hiroto ^{[2
]}

Kawachi, Tomoyuki ^{[1
]}

Kume, Shoen ^{[3
]}

Takahashi, Koji ^{[2
]}

机构：

[1] SHIONOGI & CO LTD, Drug Metab & Pharmacokinet, Res Lab Dev, 3-1-1 Toyonaka, Osaka 5610825, Japan

[2] SHIONOGI & CO LTD, Drug Discovery Technol, Drug Discovery & Dis Res Lab, 3-1-1 Toyonaka, Osaka 5610825, Japan

[3] Tokyo Inst Technol, Sch Life Sci & Technol, Dept Life Sci & Technol, Yokohama, Kanagawa 2268501, Japan

来源：

SCIENTIFIC REPORTS | 2020年 / 10卷 / 01期

关键词：

IN-VITRO; PREDICTION; PERMEABILITY; ABSORPTION; DIFFERENTIATION; MEMBRANE; SYSTEM; MODEL;

D O I：

10.1038/s41598-020-63151-z

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Drug absorption via the intestinal tissue is modulated by membrane permeability and metabolism in intestinal epithelial cells (IECs). In drug discovery research, using human IECs to evaluate membrane permeability and metabolic stability can offer very useful information when exploring for drug candidate compounds that have good bioavailability and when trying to predict the fraction absorbed and intestinal availability in humans. Here, we evaluated the pharmacokinetic functions of human IECs differentiated from human induced pluripotent stem cells (hiPSCs) in 3D cultures. As human IECs differentiated in 3D cultures form intestinal organoids and spheroids (herein termed organoids), their morphology makes it difficult to evaluate their pharmacokinetic functions. Therefore, we dissociated intestinal organoids into single cells and attempted to purify human IECs. We found that hiPSC-derived IECs (hiPSC-IECs) expressed the epithelial cell adhesion molecule (EpCAM) and could be highly purified by sorting EpCAM+ cells. The hiPSC-IEC monolayer showed a high TEER value (approximately 350 O x cm(2)). In addition, hiPSC-IECs oxidatively metabolized terfenadine (CYP3A and CYP2J2 substrate) and midazolam (CYP3A substrate). These results indicated that hiPSC-IECs form tight-junction and have cytochrome P450 enzymatic activities. In conclusion, we developed a novel application of hiPSC-derived intestinal organoids for drug testing.

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页数：11

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