Protocol to derive postganglionic parasympathetic neurons using human pluripotent stem cells for electrophysiological and functional assessment

Summary: Accessing untransformed human primary parasympathetic neurons is extremely challenging due to their inaccessibility in patients. Therefore, a human pluripotent stem cell-based approach to generate pure parasympathetic neurons is valuable for studying their development, maturation, and patho...

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Bibliographic Details
Main Authors: Christina James, Hsueh-Fu Wu, Jennifer Art, Archie Patel, Nadja Zeltner
Format: Article
Language:English
Published: Elsevier 2025-09-01
Series:STAR Protocols
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Online Access:http://www.sciencedirect.com/science/article/pii/S2666166725004071
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Summary:Summary: Accessing untransformed human primary parasympathetic neurons is extremely challenging due to their inaccessibility in patients. Therefore, a human pluripotent stem cell-based approach to generate pure parasympathetic neurons is valuable for studying their development, maturation, and pathology. Here, we present a protocol to derive parasympathetic neurons from human pluripotent stem cells. We describe steps for human pluripotent stem cell replating, neural crest replating, cryobanking, and parasympathetic neuron differentiation. We then detail procedures for functional assays via multielectrode array (MEA) analysis.For complete details on the use and execution of this protocol, please refer to Wu et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
ISSN:2666-1667