Generation of enterocyte-like cells from human induced pluripotent stem cells for drug absorption and metabolism studies in human small intestine.

Tatsuya Ozawa,Ryota Okamoto,Hiroyuki Mizuguchi,Ryosuke Negoro,Masashi Tachibana,Kenji Kawabata,Kazuo Takayama,Fuminori Sakurai

doi:10.1038/srep16479

Abstract

Enterocytes play an important role in drug absorption and metabolism. However, a widely used enterocyte model, Caco-2 cell, has difficulty in evaluating both drug absorption and metabolism because the expression levels of some drug absorption and metabolism-related genes in these cells differ largely from those of human enterocytes. Therefore, we decided to generate the enterocyte-like cells from human induced pluripotent stem (iPS) cells (hiPS-ELCs), which are applicable to drug absorption and metabolism studies. The efficiency of enterocyte differentiation from human iPS cells was significantly improved by using EGF, SB431542, and Wnt3A, and extending the differentiation period. The gene expression levels of cytochrome P450 3A4 (CYP3A4) and peptide transporter 1 in the hiPS-ELCs were higher than those in Caco-2 cells. In addition, CYP3A4 expression in the hiPS-ELCs was induced by treatment with 1, 25-dihydroxyvitamin D3 or rifampicin, which are known to induce CYP3A4 expression, indicating that the hiPS-ELCs have CYP3A4 induction potency. Moreover, the transendothelial electrical resistance (TEER) value of the hiPS-ELC monolayer was approximately 240 Ω*cm2, suggesting that the hiPS-ELC monolayer could form a barrier. In conclusion, we succeeded in establishing an enterocyte model from human iPS cells which have potential to be applied for drug absorption and metabolism studies.

Highlights

Enterocytes play an important role in drug absorption and metabolism
Some groups have reported that intestinal tissues and intestinal organoids, which are consist of all four intestinal cell types, could be differentiated from human pluripotent stem cells in vitro[15,16,17,18]
Many groups have demonstrated that intestinal cells could be differentiated from human pluripotent stem cells[16,19,20,21,26], a protocol for enterocyte monolayer differentiation for drug absorption and metabolism studies has not been well established, to our knowledge

Summary

Introduction

A widely used enterocyte model, Caco-2 cell, has difficulty in evaluating both drug absorption and metabolism because the expression levels of some drug absorption and metabolism-related genes in these cells differ largely from those of human enterocytes. We decided to generate the enterocytelike cells from human induced pluripotent stem (iPS) cells (hiPS-ELCs), which are applicable to drug absorption and metabolism studies. It is difficult to accurately evaluate CYP3A4-mediated drug metabolism and the CYP3A4 induction potency of drugs because the CYP3A4 expression levels in Caco-2 cells are much lower than those in the enterocytes[11,12]. We aimed to generate enterocyte-like cells from human iPS cells (hiPS-ELCs) which could evaluate both drug absorption and metabolism. We decided to generate enterocyte monolayers, which have drug absorption and metabolism capacities, from human iPS cells

Objectives

Methods

Results

Conclusion