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

Dominique C Stephens; Margaret Mungai; Amber Crabtree; Heather K Beasley; Edgar Garza-Lopez; Larry Vang; Kit Neikirk; Zer Vue; Neng Vue; Andrea G Marshall; Kyrin Turner; Jian-Qiang Shao; Bishnu Sarker; Sandra Murray; Jennifer A Gaddy; Jamaine Davis; Steven M Damo; Antentor O Hinton Jr

doi:10.1016/j.xpro.2023.102591

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

STAR Protoc. 2023 Dec 15;4(4):102591. doi: 10.1016/j.xpro.2023.102591. Epub 2023 Nov 7.

Authors

Dominique C Stephens¹, Margaret Mungai², Amber Crabtree³, Heather K Beasley³, Edgar Garza-Lopez², Larry Vang³, Kit Neikirk³, Zer Vue³, Neng Vue³, Andrea G Marshall³, Kyrin Turner⁴, Jian-Qiang Shao⁵, Bishnu Sarker⁶, Sandra Murray⁷, Jennifer A Gaddy⁸, Jamaine Davis⁹, Steven M Damo¹⁰, Antentor O Hinton Jr¹¹

Affiliations

¹ Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN 37232, USA; Department of Life and Physical Sciences, Fisk University, Nashville, TN 37232, USA.
² Department of Internal Medicine, University of Iowa, Iowa City, IA 52242, USA.
³ Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN 37232, USA.
⁴ Department of Life and Physical Sciences, Fisk University, Nashville, TN 37232, USA.
⁵ Central Microscopy Research Facility, University of Iowa, Iowa City, IA 52242, USA.
⁶ School of Applied Computational Sciences, Meharry Medical College, Nashville, TN 37232, USA.
⁷ Department of Cell Biology, College of Medicine, University of Pittsburgh, Pittsburgh, TN 15260, USA.
⁸ 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.
⁹ Department of Biochemistry and Cancer Biology. Meharry Medical College, Nashville, TN, USA.
¹⁰ Department of Life and Physical Sciences, Fisk University, Nashville, TN 37232, USA. Electronic address: [email protected].
¹¹ Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN 37232, USA. Electronic address: [email protected].

Abstract

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.

Keywords: Cell Culture; Cell Isolation; Model Organisms.

MeSH terms

Animals
Cell Differentiation / physiology
Fluorescent Antibody Technique
Mice
Muscle Fibers, Skeletal*
Muscle, Skeletal*
Myoblasts

Abstract

MeSH terms

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