Generation of Human-Induced Pluripotent Stem Cell-Derived Functional Enterocyte-Like Cells for Pharmacokinetic Studies

• Published in: Stem cell reports Vol. 16; no. 2; pp. 295 - 308
• Main Authors: Yoshida, Shinpei, Honjo, Takayuki, Iino, Keita, Ishibe, Ryunosuke, Leo, Sylvia, Shimada, Tomoka, Watanabe, Teruhiko, Ishikawa, Masaya, Maeda, Kazuya, Kusuhara, Hiroyuki, Shiraki, Nobuaki, Kume, Shoen
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 09.02.2021; Elsevier
• Subjects: Adult; ATP Binding Cassette Transporter, Subfamily G, Member 2 - metabolism; Cell Culture Techniques - methods; Cell Differentiation; Cell Line; Cells, Cultured; Collagen - metabolism; Culture Media; Cytochrome P-450 CYP3A - metabolism; drug development; enterocyte; Enterocytes - cytology; Enterocytes - metabolism; Female; human model; Humans; induced pluripotent stem cells; Induced Pluripotent Stem Cells - cytology; Induced Pluripotent Stem Cells - metabolism; intestine; Intestine, Small - cytology; Intestine, Small - metabolism; in vitro differentiation; Male; Middle Aged; Neoplasm Proteins - metabolism; pharmacokinetics; Young Adult; intestine; in vitro differentiation; drug development; pharmacokinetics; induced pluripotent stem cells; enterocyte; human model
• ISSN: 2213-6711; 2213-6711
• DOI: 10.1016/j.stemcr.2020.12.017

We aimed to establish an in vitro differentiation procedure to generate matured small intestinal cells mimicking human small intestine from human-induced pluripotent stem cells (iPSCs). We previously reported the efficient generation of CDX2-expressing intestinal progenitor cells from embryonic stem cells (ESCs) using 6-bromoindirubin-3′-oxime (BIO) and (3,5-difluorophenylacetyl)-L-alanyl-L-2-phenylglycine tert-butyl ester (DAPT) to treat definitive endodermal cells. Here, we demonstrate the generation of enterocyte-like cells by culturing human iPSC-derived intestinal progenitor cells on a collagen vitrigel membrane (CVM) and treating cells with a simple maturation medium containing BIO, DMSO, dexamethasone, and activated vitamin D3. Functional tests further confirmed that these iPSC-derived enterocyte-like cells exhibit P-gp- and BCRP-mediated efflux and cytochrome P450 3A4 (CYP3A4)-mediated metabolism. We concluded that hiPS cell-derived enterocyte-like cells can be used as a model for the evaluation of drug transport and metabolism studies in the human small intestine. •hiPSC-derived intestinal progenitors are differentiated into enterocytes•The collagen vitrigel membrane supports hiPSCs to mature into functional enterocytes•iPSC-derived enterocytes show efflux transporter and metabolizing enzyme activities•iPSC-derived enterocytes can be used as a human small intestine model We established a culture procedure to generate hiPSC-derived enterocyte-like cells that can be used as a model for the evaluation of drug transport and metabolism studies in the human small intestine.