Differentiation of human pluripotent stem cells (hPSCs) into organ-specific subtypes offers an exciting avenue for the study of embryonic development and disease processes, for pharmacologic studies and as a potential resource for therapeutic transplant1,2. To date, limited in vivo models exist for human intestine, all of which are dependent upon primary epithelial cultures or digested tissue from surgical biopsies that include mesenchymal cells transplanted on biodegradable scaffolds3,4. Here, we generated human intestinal organoids (HIOs) produced in vitro from human embryonic stem cells (ESCs) or induced pluripotent stem cells (iPSCs)5,6 that can engraft in vivo. These HIOs form mature human intestinal epithelium with intestinal stem cells contributing to the crypt-villus architecture and a laminated human mesenchyme, both supported by mouse vasculature ingrowth. In vivo transplantation resulted in marked expansion and maturation of the epithelium and mesenchyme, as demonstrated by differentiated intestinal cell lineages (enterocytes, goblet cells, Paneth cells, tuft cells and enteroendocrine cells), presence of functional brush-border enzymes (lactase, sucrase-isomaltase and dipeptidyl peptidase 4) and visible subepithelial and smooth muscle layers when compared with HIOs in vitro. Transplanted intestinal tissues demonstrated digestive functions as shown by permeability and peptide uptake studies. Furthermore, transplanted HIO-derived tissue was responsive to systemic signals from the host mouse following ileocecal resection, suggesting a role for circulating factors in the intestinal adaptive response7,8,9. This model of the human small intestine may pave the way for studies of intestinal physiology, disease and translational studies.
This is a preview of subscription content
Subscribe to Journal
Get full journal access for 1 year
only $4.92 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Tax calculation will be finalised during checkout.
Get time limited or full article access on ReadCube.
All prices are NET prices.
Cheng, X. et al. Self-renewing endodermal progenitor lines generated from human pluripotent stem cells. Cell Stem Cell 10, 371–384 (2012).
Wells, J.M. & Spence, J.R. How to make an intestine. Development 141, 752–760 (2014).
Lahar, N. et al. Intestinal subepithelial myofibroblasts support in vitro and in vivo growth of human small intestinal epithelium. PLoS ONE 6, e26898 (2011).
Levin, D.E. et al. Human tissue-engineered small intestine forms from postnatal progenitor cells. J. Pediatr. Surg. 48, 129–137 (2013).
Spence, J.R. et al. Directed differentiation of human pluripotent stem cells into intestinal tissue in vitro. Nature 470, 105–109 (2011).
McCracken, K.W., Howell, J.C., Wells, J.M. & Spence, J.R. Generating human intestinal tissue from pluripotent stem cells in vitro. Nat. Protoc. 6, 1920–1928 (2011).
Williamson, R.C., Buchholtz, T.W. & Malt, R.A. Humoral stimulation of cell proliferation in small bowel after transection and resection in rats. Gastroenterology 75, 249–254 (1978).
Juno, R.J. et al. A serum factor after intestinal resection stimulates epidermal growth factor receptor signaling and proliferation in intestinal epithelial cells. Surgery 132, 377–383 (2002).
Juno, R.J., Knott, A.W., Erwin, C.R. & Warner, B.W. A serum factor(s) after small bowel resection induces intestinal epithelial cell proliferation: effects of timing, site, and extent of resection. J. Pediatr. Surg. 38, 868–874 (2003).
Simon-Assmann, P., Turck, N., Sidhoum-Jenny, M., Gradwohl, G. & Kedinger, M. In vitro models of intestinal epithelial cell differentiation. Cell Biol.Toxicol. 23, 241–256 (2007).
Jung, P. et al. Isolation and in vitro expansion of human colonic stem cells. Nat. Med. 17, 1225–1227 (2011).
Sato, T. et al. Long-term expansion of epithelial organoids from human colon, adenoma, adenocarcinoma, and Barrett's epithelium. Gastroenterology 141, 1762–1772 (2011).
Campbell, F.C., Tait, I.S., Flint, N. & Evans, G.S. Transplantation of cultured small bowel enterocytes. Gut 34, 1153–1155 (1993).
Agopian, V.G., Chen, D.C., Avansino, J.R. & Stelzner, M. Intestinal stem cell organoid transplantation generates neomucosa in dogs. J. Gastrointest. Surg. 13, 971–982 (2009).
Avansino, J.R., Chen, D.C., Hoagland, V.D., Woolman, J.D. & Stelzner, M. Orthotopic transplantation of intestinal mucosal organoids in rodents. Surgery 140, 423–434 (2006).
Tait, I.S., Evans, G.S., Flint, N. & Campbell, F.C. Colonic mucosal replacement by syngeneic small intestinal stem cell transplantation. Am. J. Surg. 167, 67–72 (1994).
Tait, I.S., Flint, N., Campbell, F.C. & Evans, G.S. Generation of neomucosa in vivo by transplantation of dissociated rat postnatal small intestinal epithelium. Differentiation 56, 91–100 (1994).
Fordham, R.P. et al. Transplantation of expanded fetal intestinal progenitors contributes to colon regeneration after injury. Cell Stem Cell 13,734–744 (2013).
Yui, S. et al. Functional engraftment of colon epithelium expanded in vitro from a single adult Lgr5(+) stem cell. Nat. Med. 18, 618–623 (2012).
Kosinski, C. et al. Indian hedgehog regulates intestinal stem cell fate through epithelial-mesenchymal interactions during development. Gastroenterology 139, 893–903 (2010).
McLin, V.A., Henning, S.J. & Jamrich, M. The role of the visceral mesoderm in the development of the gastrointestinal tract. Gastroenterology 136, 2074–2091 (2009).
Zorn, A.M. & Wells, J.M. Vertebrate endoderm development and organ formation. Annu. Rev. Cell Dev. Biol. 25, 221–251 (2009).
Gracz, A.D. et al. Brief report: CD24 and CD44 mark human intestinal epithelial cell populations with characteristics of active and facultative stem cells. Stem Cells 31, 2024–2030 (2013).
Kroon, E. et al. Pancreatic endoderm derived from human embryonic stem cells generates glucose-responsive insulin-secreting cells in vivo. Nat. Biotechnol. 26, 443–452 (2008).
Kovalenko, P.L. & Basson, M.D. The correlation between the expression of differentiation markers in rat small intestinal mucosa and the transcript levels of schlafen 3. JAMA Surg. 148, 1013–1019 (2013).
Dekaney, C.M. et al. Expansion of intestinal stem cells associated with long-term adaptation following ileocecal resection in mice. Am. J. Physiol. Gastrointest. Liver Physiol. 293, G1013–G1022 (2007).
D'Amour, K.A. et al. Efficient differentiation of human embryonic stem cells to definitive endoderm. Nat. Biotechnol. 23, 1534–1541 (2005).
Warlich, E. et al. Lentiviral vector design and imaging approaches to visualize the early stages of cellular reprogramming. Mol. Ther. 19, 782–789 (2011).
Wang, F. et al. Isolation and characterization of intestinal stem cells based on surface marker combinations and colony-formation assay. Gastroenterology 145, 383–395.e1–e21 (2013).
Groneberg, D.A. et al. Intestinal peptide transport: ex vivo uptake studies and localization of peptide carrier PEPT1. Am. J. Physiol. Gastrointest. Liver Physiol. 281, G697–G704 (2001).
This project was supported in part by US National Institutes of Health (NIH) grants NIH-DK092456 (J.M.W. and N.F.S.), NIH-U18NS080815 and R01DK098350 (J.M.W.), NIH-DK092306 and NIH-CA142826 (N.F.S.), NIH-R01DK083325 (M.A.H.), NIH P30 DK078392 (Digestive Health Center, Cincinnati Children Hospital Medical Center), NIH UL1RR026314 (Clinical and Translational Science Awards (CTSA), University of Cincinnati), NIH-DK36729 (G.G.), NIH-K01DK091415 (J.R.S.), NIH-P30DK034933 (University of Michigan) and NIH-DK094775 (S.R.F.).
The authors declare no competing financial interests.
About this article
Cite this article
Watson, C., Mahe, M., Múnera, J. et al. An in vivo model of human small intestine using pluripotent stem cells. Nat Med 20, 1310–1314 (2014). https://doi.org/10.1038/nm.3737
Journal of Biological Engineering (2022)
Signal Transduction and Targeted Therapy (2022)
Nature Reviews Gastroenterology & Hepatology (2022)
Tissue extracellular matrix hydrogels as alternatives to Matrigel for culturing gastrointestinal organoids
Nature Communications (2022)
Nature Reviews Gastroenterology & Hepatology (2022)