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Analyzing Mitochondrial Function in Brown Adipocytes with a Bioenergetic Analyzer

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Book cover Clinical and Preclinical Models for Maximizing Healthspan

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2138))

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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Author notes

  1. Justin Darcy and Chih-Hao Wang contributed equally to this work.

Authors and Affiliations

  1. Section on Integrative Physiology and Metabolism, Joslin Diabetes Center, Harvard Medical School, Boston, MA, USA

    Justin Darcy, Chih-Hao Wang & Yu-Hua Tseng

  2. Harvard Stem Cell Institute, Harvard University, Cambridge, MA, USA

    Yu-Hua Tseng

Authors
  1. Justin Darcy
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  2. Chih-Hao Wang
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  3. Yu-Hua Tseng
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Corresponding author

Correspondence to Yu-Hua Tseng .

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Editors and Affiliations

  1. Laboratory of Neuroproteomics, Department of Biochemistry and Tissue Biology, Institute of Biology, University of Campinas (UNICAMP), Campinas, São Paulo, Brazil

    Paul C. Guest

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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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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0470-0

  • Online ISBN: 978-1-0716-0471-7

  • eBook Packages: Springer Protocols

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