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

By J. Spence, C. Mayhew, S. Rankin, M. Kuhar, Jefferson E. Vallance, Kathryn Tolle, Elizabeth E. Hoskins, V. Kalinichenko, S. Wells, Aaron M. Zorn and 2 more. Nature, 2010

1,804 citations recorded by the source corpus.

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.

Subfields: Hepatology, Molecular Biology, Surgery

View Paper: https://www.semanticscholar.org/paper/29bfc98c9942327fa3ab51db1043595569e83007

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Citations in the corpus

Organoid modeling of the tumor immune microenvironment (2018, background): “T cells, iPSC-based organoids with stromal elements (Spence et al., 2011), or mediation of anti-PD-1 effects by TAMs (Gordon et al.”
Stem Cell Organoid Engineering (2016, neutral): “, 2013), intestine (Spence et al., 2011), liver (Takebe et al.”
Wnt/ b -Catenin Signaling, Disease, and Emerging Therapeutic Modalities (2017, background): “Similar cultures that additionally contained mesenchymal elements could be started from induced pluripotent stem cells (iPSCs) (Spence et al., 2011) This culture systemhas since been adapted to grow organoids from Wnt-dependent adult stem cells from the epithelial compartments of a growing number of”
In vitro generation of human pluripotent stem cell derived lung organoids (2015, methodology): “…of FGF and WNT signaling synergistically drives CDX2+ intestinal lineage commitment in hPSC-derived endoderm and also drives ‘morphogenesis in a dish’, where the 2D tissues self-organize into 3D spheroids comprised of mesenchymal and polarized epithelial layers that detach from the adherent…”