Click here to close Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly. We suggest using a current version of Chrome, FireFox, or Safari.
Nature. February 3, 2011; 470 (7332): 105-9.

Directed differentiation of human pluripotent stem cells into intestinal tissue in vitro.

Spence JR , Mayhew CN , Rankin SA , Kuhar MF , Vallance JE , Tolle K , Hoskins EE , Kalinichenko VV , Wells SI , Zorn AM , Shroyer NF , Wells JM .

Studies in embryonic development have guided successful efforts to direct the differentiation of human embryonic and induced pluripotent stem cells (PSCs) into specific organ cell types in vitro. For example, human PSCs have been differentiated into monolayer cultures of liver hepatocytes and pancreatic endocrine cells that have therapeutic efficacy in animal models of liver disease and diabetes, respectively. However, the generation of complex three-dimensional organ tissues in vitro remains a major challenge for translational studies. Here we establish a robust and efficient process to direct the differentiation of human PSCs into intestinal tissue in vitro using a temporal series of growth factor manipulations to mimic embryonic intestinal development. This involved activin-induced definitive endoderm formation, FGF/Wnt-induced posterior endoderm pattering, hindgut specification and morphogenesis, and a pro-intestinal culture system to promote intestinal growth, morphogenesis and cytodifferentiation. The resulting three-dimensional intestinal ''organoids'' consisted of a polarized, columnar epithelium that was patterned into villus-like structures and crypt-like proliferative zones that expressed intestinal stem cell markers. The epithelium contained functional enterocytes, as well as goblet, Paneth and enteroendocrine cells. Using this culture system as a model to study human intestinal development, we identified that the combined activity of WNT3A and FGF4 is required for hindgut specification whereas FGF4 alone is sufficient to promote hindgut morphogenesis. Our data indicate that human intestinal stem cells form de novo during development. We also determined that NEUROG3, a pro-endocrine transcription factor that is mutated in enteric anendocrinosis, is both necessary and sufficient for human enteroendocrine cell development in vitro. PSC-derived human intestinal tissue should allow for unprecedented studies of human intestinal development and disease.

PubMed ID: 21151107
PMC ID: PMC3033971
Article link: Nature.
Grant support: F32 DK83202-01 NIDDK NIH HHS , P30 DK078392 NIDDK NIH HHS , R01 CA142826 NCI NIH HHS , R01 CA142826-02 NCI NIH HHS , R01DK080823A1 NIDDK NIH HHS , R01GM072915 NIGMS NIH HHS , R03 DK084167 NIDDK NIH HHS , R03 DK084167-02 NIDDK NIH HHS , T32 HD07463 NICHD NIH HHS , U54 RR025216 NCRR NIH HHS

Genes referenced: ascl2 cdh1 cdx2 cga chga fgf4 klf5 lgr5 muc2 neurog3 npat slc7a5 sox9 wnt3a

External Resources:
Article Images: [+] show captions

Baker, 2011, Pubmed[+]

Xenbase: The Xenopus laevis and X. tropicalis resource.
Version: 4.9.0
Major funding for Xenbase is provided by the National Institute of Child Health and Human Development, grant P41 HD064556