A xenogeneic-free system generating functional human gut organoids from pluripotent stem cells

JCI Insight. 2017 Jan 12;2(1):e86492. doi: 10.1172/jci.insight.86492.


Functional intestines are composed of cell types from all 3 primary germ layers and are generated through a highly orchestrated and serial developmental process. Directed differentiation of human pluripotent stem cells (hPSCs) has been shown to yield gut-specific cell types; however, these structures do not reproduce critical functional interactions between cell types of different germ layers. Here, we developed a simple protocol for the generation of mature functional intestinal organoids from hPSCs under xenogeneic-free conditions. The stem cell-derived gut organoids produced here were found to contain distinct types of intestinal cells, including enterocytes, goblet cells, Paneth cells, and enteroendocrine cells, that were derived from all 3 germ layers; moreover, they demonstrated intestinal functions, including peptide absorption, and showed innervated bowel movements in response to stimulation with histamine and anticholinergic drugs. Importantly, the gut organoids obtained using this xenogeneic-free system could be stably maintained in culture for prolonged periods and were successfully engrafted in vivo. Our xenogeneic-free approach for generating gut organoids from hPSCs provides a platform for studying human intestinal diseases and for pharmacological testing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Differentiation / genetics
  • Enterocytes / cytology
  • Enterocytes / metabolism
  • Goblet Cells / cytology
  • Goblet Cells / metabolism
  • Humans
  • Induced Pluripotent Stem Cells / cytology*
  • Induced Pluripotent Stem Cells / metabolism
  • Induced Pluripotent Stem Cells / transplantation
  • Intestines / cytology*
  • Octamer Transcription Factor-3
  • Organoids / cytology*
  • Organoids / metabolism
  • Paneth Cells / cytology
  • Paneth Cells / metabolism
  • Pluripotent Stem Cells / cytology*
  • SOXB1 Transcription Factors


  • Octamer Transcription Factor-3
  • POU5F1 protein, human
  • SOX1 protein, human
  • SOXB1 Transcription Factors