In vitro patterning of pluripotent stem cell-derived intestine recapitulates in vivo human development

Development. 2017 Mar 15;144(6):1045-1055. doi: 10.1242/dev.138453. Epub 2016 Dec 7.

Abstract

The intestine plays a central role in digestion, nutrient absorption and metabolism, with individual regions of the intestine having distinct functional roles. Many examples of region-specific gene expression in the adult intestine are known, but how intestinal regional identity is established during development is a largely unresolved issue. Here, we have identified several genes that are expressed in a region-specific manner in the developing human intestine. Using human embryonic stem cell-derived intestinal organoids, we demonstrate that the duration of exposure to active FGF and WNT signaling controls regional identity. Short-term exposure to FGF4 and CHIR99021 (a GSK3β inhibitor that stabilizes β-catenin) resulted in organoids with gene expression patterns similar to developing human duodenum, whereas longer exposure resulted in organoids similar to ileum. When region-specific organoids were transplanted into immunocompromised mice, duodenum-like organoids and ileum-like organoids retained their regional identity, demonstrating that regional identity of organoids is stable after initial patterning occurs. This work provides insights into the mechanisms that control regional specification of the developing human intestine and provides new tools for basic and translational research.

Keywords: Human; Intestine; Organoid; Patterning; Pluripotent stem cells.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biomarkers / metabolism
  • Body Patterning* / genetics
  • Cell Differentiation / genetics
  • Computational Biology
  • Embryonic Development* / genetics
  • Fetus / embryology*
  • Fibroblast Growth Factors / metabolism
  • Gene Expression Profiling
  • Humans
  • Intestines / embryology*
  • Mice
  • Organoids / metabolism
  • Organoids / transplantation
  • Pluripotent Stem Cells / cytology*
  • Pluripotent Stem Cells / metabolism
  • Reproducibility of Results
  • Sequence Analysis, RNA
  • Signal Transduction / genetics
  • Wnt Proteins / metabolism

Substances

  • Biomarkers
  • Wnt Proteins
  • Fibroblast Growth Factors