Protocol for generating postganglionic sympathetic neurons using human pluripotent stem cells for electrophysiological and functional assessments

Hsueh-Fu Wu; Jennifer Art; Tripti Saini; Nadja Zeltner

STAR Protocols (Jun 2024)

Protocol for generating postganglionic sympathetic neurons using human pluripotent stem cells for electrophysiological and functional assessments

Hsueh-Fu Wu,
Jennifer Art,
Tripti Saini,
Nadja Zeltner

Affiliations

Hsueh-Fu Wu: Center for Molecular Medicine, University of Georgia, Athens, GA 30602, USA; Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA 30602, USA
Jennifer Art: Center for Molecular Medicine, University of Georgia, Athens, GA 30602, USA; Biomedical and Translational Sciences Institute, Neuroscience Program, University of Georgia, Athens, GA 30602, USA
Tripti Saini: Center for Molecular Medicine, University of Georgia, Athens, GA 30602, USA; Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA 30602, USA
Nadja Zeltner: Center for Molecular Medicine, University of Georgia, Athens, GA 30602, USA; Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA 30602, USA; Department of Cellular Biology, University of Georgia, Athens, GA 30602, USA; Corresponding author

Journal volume & issue: Vol. 5, no. 2
p. 102970

Abstract

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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.

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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