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An in vivo model of human small intestine using pluripotent stem cells

Abstract

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.

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Figure 1: HIOs engraft in vivo to form mature intestinal tissue.
Figure 2: Engrafted intestinal tissue resembles adult intestine and is almost entirely of human origin.
Figure 3: Engrafted tissue matures in vivo and resembles mature small intestine.
Figure 4: Engrafted human intestinal tissue responds to humoral factors following ileocecal resection (ICR) in the mouse host.

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Acknowledgements

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.).

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Authors

Contributions

C.L.W. and M.M.M. designed the experiments, performed the surgeries, collected and assembled data and wrote the manuscript. J.M., J.C.H., N.S., H.M.P., J.I.S., J.E.V., C.N.M., Y.S., G.G., S.R.F. and J.R.S. collected and provided study materials. N.F.S. and J.M.W. were involved in the study design and wrote and approved the manuscript. M.A.H. designed the study, wrote the manuscript and finally approved the manuscript. All authors discussed the results and commented on the manuscript.

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Correspondence to Michael A Helmrath.

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The authors declare no competing financial interests.

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Supplementary Figures 1–9 and Supplementary Tables 1 and 2 (PDF 9630 kb)

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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

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