Measuring Mitochondrial Substrate Utilization in Skeletal Muscle Stem Cells

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 1668; p. 61
• Main Authors: Ly, C Hai, Ryall, James G
• Format: Journal Article
• Language: English
• Published: United States 2017
• Subjects: Animals; Cell Differentiation - drug effects; Cell Line; Cell Proliferation - drug effects; Epoxy Compounds - pharmacology; Fatty Acids - metabolism; Glucose - metabolism; Glutamic Acid - metabolism; Hydrogen-Ion Concentration; Hypoglycemic Agents - pharmacology; Mitochondria, Muscle - drug effects; Mitochondria, Muscle - metabolism; Myoblasts, Skeletal - drug effects; Myoblasts, Skeletal - metabolism; Oxygen Consumption - drug effects; Smegmamorpha; Sulfides - pharmacology; Thiadiazoles - pharmacology; Fatty acid oxidation; Glycolysis; Glutaminolysis; Metabolism; Muscle stem cells
• ISSN: 1940-6029
• DOI: 10.1007/978-1-4939-7283-8_5

Skeletal muscle stem cells (MuSCs) derived from the somatic mesoderm play a critical role in successful muscle regeneration following injury and trauma. MuSCs have been found to undergo rapid changes in metabolism following a change in cell state, such as that which occurs during the transition from quiescence to an actively proliferating state. There is mounting evidence that metabolism is critically important in the regulation of quiescence, activation, and differentiation and thus the development of new techniques that aim to further probe the metabolism of MuSCs is essential. The Seahorse XF Bioanalyzer is a powerful tool that simultaneously measures the extracellular rate of change in oxygen partial pressure and pH, providing a method to measure mitochondrial respiration and lactate production. In this chapter, we describe the use of key metabolic inhibitors that allow for the investigation of mitochondrial substrate utilization in primary MuSCs.