Protocol for isolating mice skeletal muscle myoblasts and myotubes via differential antibody validation.

Cell Culture Cell Isolation Model Organisms

Journal

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

Informations de publication

Date de publication:
07 Nov 2023
Historique:
received: 22 05 2023
revised: 06 08 2023
accepted: 01 09 2023
medline: 8 11 2023
pubmed: 8 11 2023
entrez: 8 11 2023
Statut: aheadofprint

Résumé

Isolation of skeletal muscles allows for the exploration of many complex diseases. Here, we present a protocol for isolating mice skeletal muscle myoblasts and myotubes that have been differentiated through antibody validation. We describe steps for collecting and preparing murine skeletal tissue, myoblast cell maintenance, plating, and cell differentiation. We then detail procedures for cell incubation, immunostaining, slide preparation and storage, and imaging for immunofluorescence validation.

Identifiants

pubmed: 37938976
pii: S2666-1667(23)00558-0
doi: 10.1016/j.xpro.2023.102591
pmc: PMC10663959
pii:
doi:

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

102591

Subventions

Organisme : NCATS NIH HHS
ID : UL1 TR000445
Pays : United States
Organisme : NICHD NIH HHS
ID : R01 HD090061
Pays : United States
Organisme : NHLBI NIH HHS
ID : R25 HL106365
Pays : United States
Organisme : NCRR NIH HHS
ID : UL1 RR024975
Pays : United States
Organisme : BLRD VA
ID : I01 BX005352
Pays : United States
Organisme : NIDDK NIH HHS
ID : P30 DK020593
Pays : United States

Commentaires et corrections

Type : UpdateOf

Informations de copyright

Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.

Déclaration de conflit d'intérêts

Declaration of interests The authors declare no competing interests.

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Auteurs

Dominique C Stephens (DC)

Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN 37232, USA; Department of Life and Physical Sciences, Fisk University, Nashville, TN 37232, USA.

Margaret Mungai (M)

Department of Internal Medicine, University of Iowa, Iowa City, IA 52242, USA.

Amber Crabtree (A)

Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN 37232, USA.

Heather K Beasley (HK)

Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN 37232, USA.

Edgar Garza-Lopez (E)

Department of Internal Medicine, University of Iowa, Iowa City, IA 52242, USA.

Larry Vang (L)

Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN 37232, USA.

Kit Neikirk (K)

Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN 37232, USA.

Zer Vue (Z)

Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN 37232, USA.

Neng Vue (N)

Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN 37232, USA.

Andrea G Marshall (AG)

Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN 37232, USA.

Kyrin Turner (K)

Department of Life and Physical Sciences, Fisk University, Nashville, TN 37232, USA.

Jian-Qiang Shao (JQ)

Central Microscopy Research Facility, University of Iowa, Iowa City, IA 52242, USA.

Bishnu Sarker (B)

School of Applied Computational Sciences, Meharry Medical College, Nashville, TN 37232, USA.

Sandra Murray (S)

Department of Cell Biology, College of Medicine, University of Pittsburgh, Pittsburgh, TN 15260, USA.

Jennifer A Gaddy (JA)

Division of Infectious Diseases, Vanderbilt University School of Medicine, Nashville, TN, USA; Tennessee Valley Healthcare Systems, U.S. Department of Veterans Affairs, Nashville, TN, USA.

Jamaine Davis (J)

Department of Biochemistry and Cancer Biology. Meharry Medical College, Nashville, TN, USA.

Steven M Damo (SM)

Department of Life and Physical Sciences, Fisk University, Nashville, TN 37232, USA. Electronic address: sdamo@fisk.edu.

Antentor O Hinton (AO)

Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN 37232, USA. Electronic address: antentor.o.hinton.jr@vanderbilt.edu.

Classifications MeSH