Abstract
Brown adipocytes are a cell type with high mitochondrial content and bioenergetic capacity. A critical means to measure mitochondrial function, macromolecule fuel usage, and other important phenotypes is with a bioenergetic analyzer. Here, we describe how to isolate, culture, and differentiate brown preadipocytes into mature adipocytes. We also explain how to perform a mitochondrial (mito) stress test, using the bioenergetic analyzer. The mito stress test is able to give researchers a plethora of insights into mitochondrial function, including basal respiration, proton leak, ATP production, maximal respiration, and reserve capacity, making it a powerful tool for analyzing brown adipocytes.
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Acknowledgments
This work was supported by the US National Institutes of Health (NIH) grants R01DK077097 and R01DK102898 (to Y.-H.T.) and by P30DK036836 (to Joslin Diabetes Center’s Diabetes Research Center). Y.-H.T. was also supported by US Army Medical Research Grant W81XWH-17-1-0428. J.D. was supported by institutional research training grant T32DK007260. C.-H.W. was supported by a Postdoctoral Research Abroad Program (PRAP) from the Ministry of Science and Technology, Taiwan (106-2917-I-564-069). Justin Darcy and Chih-Hao Wang contributed equally to this work.
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Darcy, J., Wang, CH., Tseng, YH. (2020). Analyzing Mitochondrial Function in Brown Adipocytes with a Bioenergetic Analyzer. In: Guest, P. (eds) Clinical and Preclinical Models for Maximizing Healthspan. Methods in Molecular Biology, vol 2138. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0471-7_20
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DOI: https://doi.org/10.1007/978-1-0716-0471-7_20
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