Skip to main content

Measuring Mitochondrial Uncoupling in Mouse Primary Brown Adipocytes Differentiated Ex Vivo

Part of the Methods in Molecular Biology book series (MIMB,volume 2662)


Measuring the mitochondrial respiratory capacity of brown adipocytes ex vivo is an essential approach to understand the cell-autonomous regulators of mitochondrial uncoupling in brown adipose tissue. Here, we describe two protocols to isolate brown preadipocytes from mice, their ex vivo differentiation to mature brown adipocytes and the quantification of their mitochondrial uncoupling capacity by respirometry.

Key words

This is a preview of subscription content, log in via an institution.

Buying options

USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
USD   139.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Hardcover Book
USD   179.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more


  1. Rolfe DF, Brown GC (1997) Cellular energy utilization and molecular origin of standard metabolic rate in mammals. Physiol Rev 77:731–758

    Article  CAS  PubMed  Google Scholar 

  2. Veliova M, Ferreira CM, Benador IY et al (2020) Blocking mitochondrial pyruvate import in brown adipocytes induces energy wasting via lipid cycling. EMBO Rep 21:e49634

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Wikstrom JD, Mahdaviani K, Liesa M et al (2014) Hormone-induced mitochondrial fission is utilized by brown adipocytes as an amplification pathway for energy expenditure. EMBO J 33:418–436

    CAS  PubMed  PubMed Central  Google Scholar 

Download references


We thank Barbara Cannon, Jan Nedergaard, Jakob Wikstrom, Kiana Mahdaviani, Ilan Benador, and Marcus Fernandes de Oliveira for their mentoring on the protocol using 3–4-week-old mice.

Author information

Authors and Affiliations


Corresponding author

Correspondence to Marc Liesa .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Veliova, M., Tol, M.J., Tontonoz, P., Shirihai, O.S., Liesa, M. (2023). Measuring Mitochondrial Uncoupling in Mouse Primary Brown Adipocytes Differentiated Ex Vivo. In: Lodhi, I.J. (eds) Thermogenic Fat. Methods in Molecular Biology, vol 2662. Humana, New York, NY.

Download citation

  • DOI:

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3166-9

  • Online ISBN: 978-1-0716-3167-6

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics