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Related Experiment Video

Updated: Feb 24, 2026

Measuring Mitochondrial Substrate Flux in Recombinant Perfringolysin O-Permeabilized Cells

Measuring Mitochondrial Substrate Flux in Recombinant Perfringolysin O-Permeabilized Cells

Published on: August 13, 2021

Measuring Mitochondrial Substrate Utilization in Skeletal Muscle Stem Cells.

C Hai Ly¹, James G Ryall²

¹Stem Cell Metabolism and Regenerative Medicine Group, Basic and Clinical Myology Laboratory, The University of Melbourne, Parkville, VIC, 3010, Australia.

Methods in Molecular Biology (Clifton, N.J.)

|August 27, 2017

Summary

Investigating skeletal muscle stem cell (MuSC) metabolism is crucial for understanding muscle regeneration. New techniques using metabolic inhibitors with the Seahorse XF Bioanalyzer reveal insights into MuSC mitochondrial function.

Keywords:

Fatty acid oxidation Glutaminolysis Glycolysis Metabolism Muscle stem cells

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Last Updated: Feb 24, 2026

Measuring Mitochondrial Substrate Flux in Recombinant Perfringolysin O-Permeabilized Cells

Measuring Mitochondrial Substrate Flux in Recombinant Perfringolysin O-Permeabilized Cells

Published on: August 13, 2021

Using Isolated Mitochondria from Minimal Quantities of Mouse Skeletal Muscle for High throughput Microplate Respiratory Measurements

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Published on: October 30, 2015

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Published on: October 4, 2024

Area of Science:

Muscle stem cell biology
Cellular metabolism
Biochemistry

Background:

Skeletal muscle stem cells (MuSCs) are vital for muscle repair after injury.
MuSC metabolism shifts significantly during transitions between quiescence and proliferation.
Metabolic regulation is key to MuSC function, including activation and differentiation.

Purpose of the Study:

To highlight the importance of probing MuSC metabolism.
To describe a method for investigating mitochondrial substrate utilization in primary MuSCs.

Main Methods:

Utilizing the Seahorse XF Bioanalyzer to measure extracellular oxygen and pH changes.
Employing key metabolic inhibitors to analyze mitochondrial respiration and lactate production.
Focusing on primary MuSCs to study their metabolic profiles.

Main Results:

The Seahorse XF Bioanalyzer allows simultaneous measurement of mitochondrial respiration and glycolysis.
Metabolic inhibitors enable detailed investigation of substrate utilization pathways.
This approach provides a powerful method to characterize MuSC metabolic states.

Conclusions:

Understanding MuSC metabolism is essential for advancing muscle regeneration research.
The described methodology offers a robust platform for metabolic analysis of MuSCs.
Further investigation into MuSC metabolism can inform therapeutic strategies for muscle disorders.