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Protocol for measuring mitochondrial respiratory capacity from buccal cell swabs
Tina R Ram1, Chunlong Mu2, Jaclyn C Campbell2
1Hotchkiss Brain Institute, Cumming School of Medicine, University of Calgary, Calgary, AB T2N 4N1, Canada; Department of Biochemistry and Molecular Biology, Cumming School of Medicine, University of Calgary, Calgary, AB T2N 1N4, Canada.
STAR Protocols
|August 3, 2025
Summary
This study introduces a new, non-invasive method to measure mitochondrial respiration using cultured buccal cells. This protocol enables reliable metabolic investigation from easily accessible samples.
Area of Science:
- Biochemistry
- Cell Biology
- Metabolic Research
Background:
- Mitochondrial respirometry is crucial for assessing cellular energy production.
- Current methods often require invasive tissue biopsies, limiting accessibility.
- Non-invasive techniques for evaluating mitochondrial function are needed.
Purpose of the Study:
- To develop and validate a protocol for measuring mitochondrial respiration in cultured human buccal cells.
- To establish a reliable, non-invasive method for assessing mitochondrial function.
- To provide a tool for broader metabolic investigations.
Main Methods:
- Collection and primary culture of human buccal cells.
- Calibration of respirometry equipment.
- Measurement of oxygen consumption rates (OCR) in cultured cells.
- Cell counting for normalization of OCR data.
Main Results:
- Successful establishment of a protocol for culturing buccal cells suitable for respirometry.
- Demonstration of reliable measurement of mitochondrial oxygen consumption in these cells.
- Normalization of respiration data by cell count allows for quantitative analysis.
Conclusions:
- This protocol offers a non-invasive and accessible method for evaluating mitochondrial function.
- Cultured buccal cells are a viable sample source for mitochondrial respirometry.
- This technique provides a valuable tool for metabolic research and potentially clinical applications.
