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Protocol for generating postganglionic sympathetic neurons using human pluripotent stem cells for electrophysiological and functional assessments
oleh: Hsueh-Fu Wu, Jennifer Art, Tripti Saini, Nadja Zeltner
Format: | Article |
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Diterbitkan: | Elsevier 2024-06-01 |
Deskripsi
Summary: Assessing the development and function of the sympathetic nervous system in diseases on a large scale is challenging. Here, we present a protocol to generate human pluripotent stem cell (hPSC)-derived postganglionic sympathetic neurons (symNs) differentiated via neural crest cells (NCCs), which can be cryopreserved. We describe steps for hPSC replating, NCC replating and cryobanking, and symN differentiation. We then demonstrate the functionality of the hPSC-derived symNs, focusing on electrophysiological activity, calcium flux, and norepinephrine dynamics.For complete details on the use and execution of this protocol, please refer to Wu et al.1,2 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.