Differentiation of Enteric Nervous System Lineages from Human Pluripotent Stem Cells.

J Vis Exp

Department of Cellular and Molecular Pharmacology, University of California, San Francisco; Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California, San Francisco; Program in Craniofacial Biology, University of California, San Francisco;

Published: May 2024

The human enteric nervous system, ENS, is a large network of glial and neuronal cell types with remarkable neurotransmitter diversity. The ENS controls bowel motility, enzyme secretion, and nutrient absorption and interacts with the immune system and the gut microbiome. Consequently, developmental and acquired defects of the ENS are responsible for many human diseases and may contribute to symptoms of Parkinson's disease. Limitations in animal model systems and access to primary tissue pose significant experimental challenges in studies of the human ENS. Here, a detailed protocol is presented for effective in vitro derivation of the ENS lineages from human pluripotent stem cells, hPSC, using defined culture conditions. Our protocol begins with directed differentiation of hPSCs to enteric neural crest cells within 15 days and yields diverse subtypes of functional enteric neurons within 30 days. This platform provides a scalable resource for developmental studies, disease modeling, drug discovery, and regenerative applications.

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Source
http://dx.doi.org/10.3791/66133DOI Listing

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