Differentiation of human induced pluripotent stem cells into functional enterocyte-like cells using a simple method

Takahiro Iwao, Masashi Toyota, Yoshitaka Miyagawa, Hajime Okita, Nobutaka Kiyokawa, Hidenori Akutsu, Akihiro Umezawa, Kiyoshi Nagata, Tamihide Matsunaga

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

Human induced pluripotent stem (iPS) cells were differentiated into the endoderm using activin A and were then treated with fibroblast growth factor 2 (FGF2) for differentiation into intestinal stem cell-like cells. These immature cells were then differentiated into enterocyte-like cells using epidermal growth factor (EGF) in 2% fetal bovine serum (FBS). At the early stage of differentiation, mRNA expression of caudal type homeobox 2 (CDX2), a major transcription factor related to intestinal development and differentiation, and leucine-rich repeat-containing G-protein-coupled receptor 5 (LGR5), an intestinal stem cell marker, was markedly increased by treatment with FGF2. When cells were cultured in medium containing EGF and a low concentration of FBS, mRNAs of specific markers of intestinal epithelial cells, including sucrase-isomaltase, the intestinal oligopeptide transporter SLC15A1/peptide transporter 1 (PEPT1), and the major metabolizing enzyme CYP3A4, were expressed. In addition, sucraseisomaltase protein expression and uptake of β-Ala-Lys-N-7-amino-4-methylcoumarin-3-acetic acid (β-Ala-Lys-AMCA), a fluorescence-labeled substrate of the oligopeptide transporter, were detected. These results demonstrate a simple and direct method for differentiating human iPS cells into functional enterocyte-like cells.

Original languageEnglish
Pages (from-to)44-51
Number of pages8
JournalDrug Metabolism And Pharmacokinetics
Volume29
Issue number1
DOIs
Publication statusPublished - 2014
Externally publishedYes

Keywords

  • Drug metabolizing enzymes
  • Drug transporters
  • Enterocytes
  • Human iPS cells
  • Intestinal differentiation
  • Pharmacokinetics

ASJC Scopus subject areas

  • Pharmacology
  • Pharmaceutical Science
  • Pharmacology (medical)

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