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

31 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

Fingerprint

Induced Pluripotent Stem Cells
Enterocytes
Oligopeptides
Fibroblast Growth Factor 2
Epidermal Growth Factor
Stem Cells
Oligo-1,6-Glucosidase
Sucrase
Cytochrome P-450 CYP3A
Messenger RNA
Endoderm
Homeobox Genes
G-Protein-Coupled Receptors
Serum
Leucine
Cultured Cells
Transcription Factors
Fluorescence
Epithelial Cells
Enzymes

Keywords

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

ASJC Scopus subject areas

  • Pharmacology (medical)
  • Pharmacology
  • Pharmaceutical Science

Cite this

Differentiation of human induced pluripotent stem cells into functional enterocyte-like cells using a simple method. / Iwao, Takahiro; Toyota, Masashi; Miyagawa, Yoshitaka; Okita, Hajime; Kiyokawa, Nobutaka; Akutsu, Hidenori; Umezawa, Akihiro; Nagata, Kiyoshi; Matsunaga, Tamihide.

In: Drug Metabolism and Pharmacokinetics, Vol. 29, No. 1, 2014, p. 44-51.

Research output: Contribution to journalArticle

Iwao, Takahiro ; Toyota, Masashi ; Miyagawa, Yoshitaka ; Okita, Hajime ; Kiyokawa, Nobutaka ; Akutsu, Hidenori ; Umezawa, Akihiro ; Nagata, Kiyoshi ; Matsunaga, Tamihide. / Differentiation of human induced pluripotent stem cells into functional enterocyte-like cells using a simple method. In: Drug Metabolism and Pharmacokinetics. 2014 ; Vol. 29, No. 1. pp. 44-51.
@article{dbca0304863b468983569d132c061ab9,
title = "Differentiation of human induced pluripotent stem cells into functional enterocyte-like cells using a simple method",
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.",
keywords = "Drug metabolizing enzymes, Drug transporters, Enterocytes, Human iPS cells, Intestinal differentiation, Pharmacokinetics",
author = "Takahiro Iwao and Masashi Toyota and Yoshitaka Miyagawa and Hajime Okita and Nobutaka Kiyokawa and Hidenori Akutsu and Akihiro Umezawa and Kiyoshi Nagata and Tamihide Matsunaga",
year = "2014",
doi = "10.2133/dmpk.DMPK-13-RG-005",
language = "English",
volume = "29",
pages = "44--51",
journal = "Drug Metabolism and Pharmacokinetics",
issn = "1347-4367",
publisher = "Japanese Society for the Study of Xenobiotics",
number = "1",

}

TY - JOUR

T1 - Differentiation of human induced pluripotent stem cells into functional enterocyte-like cells using a simple method

AU - Iwao, Takahiro

AU - Toyota, Masashi

AU - Miyagawa, Yoshitaka

AU - Okita, Hajime

AU - Kiyokawa, Nobutaka

AU - Akutsu, Hidenori

AU - Umezawa, Akihiro

AU - Nagata, Kiyoshi

AU - Matsunaga, Tamihide

PY - 2014

Y1 - 2014

N2 - 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.

AB - 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.

KW - Drug metabolizing enzymes

KW - Drug transporters

KW - Enterocytes

KW - Human iPS cells

KW - Intestinal differentiation

KW - Pharmacokinetics

UR - http://www.scopus.com/inward/record.url?scp=84898488352&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84898488352&partnerID=8YFLogxK

U2 - 10.2133/dmpk.DMPK-13-RG-005

DO - 10.2133/dmpk.DMPK-13-RG-005

M3 - Article

C2 - 23822979

AN - SCOPUS:84898488352

VL - 29

SP - 44

EP - 51

JO - Drug Metabolism and Pharmacokinetics

JF - Drug Metabolism and Pharmacokinetics

SN - 1347-4367

IS - 1

ER -