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

Biotechnology and bioengineering Cell Biology Developmental biology Neuroscience Stem Cells

Journal

STAR protocols
ISSN: 2666-1667
Titre abrégé: STAR Protoc
Pays: United States
ID NLM: 101769501

Informations de publication

Date de publication:
21 Mar 2024
Historique:
received: 19 12 2023
revised: 14 02 2024
accepted: 05 03 2024
medline: 22 3 2024
pubmed: 22 3 2024
entrez: 22 3 2024
Statut: aheadofprint

Résumé

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.

Identifiants

DOI: 10.1016/j.xpro.2024.102970 PMID: 38517897
pubmed: 38517897
pii: S2666-1667(24)00135-7
doi: 10.1016/j.xpro.2024.102970
pii:
doi:

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

102970

Informations de copyright

Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.

Déclaration de conflit d'intérêts

Declaration of interests The authors declare no competing interests.

Auteurs

Hsueh-Fu Wu (HF)

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 (J)

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 (T)

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 (N)

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. Electronic address: nadja.zeltner@uga.edu.

Classifications MeSH