Open AccessIsolation and Differentiation of Primary Myoblasts from Mouse Skeletal Muscle Explants
Author(s) -
Megan Vaughan,
Katja Lamia
Publication year - 2019
Publication title -
journal of visualized experiments
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.596
H-Index - 91
ISSN - 1940-087X
DOI - 10.3791/60310
Subject(s) - myocyte , skeletal muscle , microbiology and biotechnology , c2c12 , biology , cellular differentiation , population , myogenesis , cell culture , explant culture , ex vivo , primary cell , in vivo , in vitro , anatomy , biochemistry , genetics , medicine , gene , environmental health
Primary myoblasts are undifferentiated proliferating precursors of skeletal muscle. They can be cultured and studied as muscle precursors or induced to differentiate into later stages of muscle development. The protocol provided here describes a robust method for the isolation and culture of a highly proliferative population of myoblast cells from young adult mouse skeletal muscle explants. These cells are useful for the study of the metabolic properties of skeletal muscle of different mouse models, as well as in other downstream applications such as transfection with exogenous DNA or transduction with viral expression vectors. The level of differentiation and metabolic profile of these cells depends on the length of exposure, and composition of the media used to induce myoblast differentiation. These methods provide a robust system for the study of mouse muscle cell metabolism ex vivo. Importantly, unlike in vivo models, the methods described here provide a cell population that can be expanded and studied with high levels of reproducibility.