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

Drug Metab Pharmacokinet. 2014;29(1):44-51. doi: 10.2133/dmpk.dmpk-13-rg-005. Epub 2013 Jul 2.


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, sucrase-isomaltase 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.

MeSH terms

  • Animals
  • Biomarkers / metabolism
  • Cattle
  • Cell Differentiation
  • Cell Line
  • Coumarins / metabolism
  • Cytochrome P-450 CYP3A / genetics
  • Cytochrome P-450 CYP3A / metabolism
  • Dipeptides / metabolism
  • Erythrocytes / cytology*
  • Erythrocytes / metabolism
  • Humans
  • Induced Pluripotent Stem Cells / cytology*
  • Induced Pluripotent Stem Cells / metabolism
  • Oligo-1,6-Glucosidase / genetics
  • Oligo-1,6-Glucosidase / metabolism
  • Peptide Transporter 1
  • RNA, Messenger / metabolism
  • Receptors, G-Protein-Coupled / metabolism
  • Sucrase / genetics
  • Sucrase / metabolism
  • Symporters / genetics
  • Symporters / metabolism


  • Biomarkers
  • Coumarins
  • Dipeptides
  • LGR5 protein, human
  • Peptide Transporter 1
  • RNA, Messenger
  • Receptors, G-Protein-Coupled
  • SLC15A1 protein, human
  • Symporters
  • alanyl-lysyl-N(epsilon)-7-amino-4-methylcoumarin-3-acetic acid
  • Cytochrome P-450 CYP3A
  • CYP3A4 protein, human
  • Oligo-1,6-Glucosidase
  • Sucrase