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A cell culture platform for quantifying metabolic substrate oxidation in bicarbonate-buffered medium
James R Krycer1, Mary Lor2, Rebecca L Fitzsimmons2
1QIMR Berghofer Medical Research Institute, Brisbane, Queensland, Australia; Faculty of Health, School of Biomedical Sciences, Queensland University of Technology, Brisbane, Queensland, Australia.
The Journal of Biological Chemistry
|December 31, 2021
Summary
This study introduces a novel bicarbonate-buffered well-plate platform for measuring cellular substrate oxidation. This high-throughput method allows for more physiological metabolic studies, aiding research into complex diseases.
Area of Science:
- Cellular metabolism
- Biochemical assays
- Disease research
Background:
- Complex diseases like cancer and diabetes involve metabolic changes.
- Measuring substrate utilization is key to understanding cellular metabolism.
- Existing high-throughput assays are incompatible with bicarbonate-buffered media.
Purpose of the Study:
- To develop a bicarbonate-buffered well-plate platform for measuring substrate oxidation.
- To enable high-throughput metabolic studies under more physiological conditions.
- To overcome limitations of previous metabolic assay platforms.
Main Methods:
- Developed a sealed well-plate environment equilibrated with carbon dioxide.
- Utilized radiolabeled carbon substrates to measure carbon dioxide production.
- Assessed metabolic flux in cultured adipocytes as a proof of principle.
Main Results:
- Successfully created a bicarbonate-buffered platform for substrate oxidation measurement.
- Demonstrated increased lipogenesis, glucose oxidation, and insulin sensitivity in adipocytes.
- Showcased the platform's compatibility with physiological bicarbonate buffering.
Conclusions:
- The new platform facilitates convenient and high-throughput study of cellular metabolism.
- Enables research under more physiologically relevant bicarbonate-buffered conditions.
- Aids in drug screening and mechanistic studies for complex diseases.
