1. Generation of human induced pluripotent stem cell-derived functional enterocyte-like cells for pharmacokinetic studies
- Author
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Hiroyuki Kusuhara, Sylvia Leo, Takayuki Honjo, Shinpei Yoshida, Ryunosuke Ishibe, Teruhiko Watanabe, Nobuaki Shiraki, Shoen Kume, Masaya Ishikawa, Tomoka Shimada, Kazuya Maeda, and Keita Iino
- Subjects
0301 basic medicine ,Adult ,Male ,Enterocyte ,induced pluripotent stem cells ,Cell Culture Techniques ,enterocyte ,Biology ,Biochemistry ,Article ,Cell Line ,03 medical and health sciences ,Young Adult ,0302 clinical medicine ,in vitro differentiation ,Intestine, Small ,Genetics ,medicine ,ATP Binding Cassette Transporter, Subfamily G, Member 2 ,Cytochrome P-450 CYP3A ,Humans ,Progenitor cell ,Induced pluripotent stem cell ,intestine ,Cells, Cultured ,CYP3A4 ,Cell Differentiation ,Cell Biology ,Metabolism ,Middle Aged ,human model ,Embryonic stem cell ,drug development ,Small intestine ,Cell biology ,Culture Media ,Neoplasm Proteins ,030104 developmental biology ,medicine.anatomical_structure ,Enterocytes ,Female ,Efflux ,Collagen ,pharmacokinetics ,030217 neurology & neurosurgery ,Developmental Biology - Abstract
Summary 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., Highlights • 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.
- Published
- 2021