Publication Date:
2010-12-15
Description:
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.〈br /〉〈br /〉〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3033971/" target="_blank"〉〈img src="https://static.pubmed.gov/portal/portal3rc.fcgi/4089621/img/3977009" border="0"〉〈/a〉 〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3033971/" target="_blank"〉This paper as free author manuscript - peer-reviewed and accepted for publication〈/a〉〈br /〉〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Spence, Jason R -- Mayhew, Christopher N -- Rankin, Scott A -- Kuhar, Matthew F -- Vallance, Jefferson E -- Tolle, Kathryn -- Hoskins, Elizabeth E -- Kalinichenko, Vladimir V -- Wells, Susanne I -- Zorn, Aaron M -- Shroyer, Noah F -- Wells, James M -- F32 DK083202/DK/NIDDK NIH HHS/ -- F32 DK083202-01/DK/NIDDK NIH HHS/ -- F32 DK83202-01/DK/NIDDK NIH HHS/ -- K01 DK091415/DK/NIDDK NIH HHS/ -- P30 DK078392/DK/NIDDK NIH HHS/ -- R01 CA142826/CA/NCI NIH HHS/ -- R01 CA142826-02/CA/NCI NIH HHS/ -- R01 DK080823/DK/NIDDK NIH HHS/ -- R01 DK080823-01A1/DK/NIDDK NIH HHS/ -- R01 DK080823-01A1S1/DK/NIDDK NIH HHS/ -- R01 DK092456/DK/NIDDK NIH HHS/ -- R01 GM072915/GM/NIGMS NIH HHS/ -- R01 GM072915-01A2/GM/NIGMS NIH HHS/ -- R01DK080823A1/DK/NIDDK NIH HHS/ -- R01GM072915/GM/NIGMS NIH HHS/ -- R03 DK084167/DK/NIDDK NIH HHS/ -- R03 DK084167-02/DK/NIDDK NIH HHS/ -- T32 HD07463/HD/NICHD NIH HHS/ -- U54 RR025216/RR/NCRR NIH HHS/ -- UL1 TR000077/TR/NCATS NIH HHS/ -- England -- Nature. 2011 Feb 3;470(7332):105-9. doi: 10.1038/nature09691. Epub 2010 Dec 12.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Division of Developmental Biology, Cincinnati Children's Hospital Medical Center, 3333 Burnet Avenue, Cincinnati, Ohio 45229-3039, USA.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/21151107" target="_blank"〉PubMed〈/a〉
Keywords:
Activins/pharmacology
;
Basic Helix-Loop-Helix Transcription Factors/genetics/metabolism
;
Body Patterning/drug effects
;
Cell Culture Techniques
;
Cell Differentiation/*drug effects
;
Cells, Cultured
;
Culture Media/chemistry/pharmacology
;
Embryonic Stem Cells/*cytology/drug effects
;
Endoderm/cytology/drug effects/embryology
;
Fibroblast Growth Factor 4/pharmacology
;
Humans
;
Induced Pluripotent Stem Cells/*cytology/drug effects
;
Intercellular Signaling Peptides and Proteins/*pharmacology
;
Intestines/anatomy & histology/*cytology/drug effects/embryology
;
Microvilli/drug effects
;
Morphogenesis/drug effects
;
Nerve Tissue Proteins/genetics/metabolism
;
Organogenesis/drug effects
;
Time Factors
;
Wnt Proteins/pharmacology
;
Wnt3 Protein
;
Wnt3A Protein
Print ISSN:
0028-0836
Electronic ISSN:
1476-4687
Topics:
Biology
,
Chemistry and Pharmacology
,
Medicine
,
Natural Sciences in General
,
Physics
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