Isolation, Culture, and Functional Analysis of Murine Thermogenic Adipocytes

Isolation, Culture, and Functional Analysis of Murine Thermogenic Adipocytes

Blog Article

Summary: Studying brown and brite adipose tissue requires precise and reliable Sealed Crimp Joiner Terminals quantification of cellular thermogenesis.This protocol describes the isolation of primary murine pre-adipocytes, differentiation into thermogenic brown and brite adipocytes, and subsequent oxygen consumption analysis.Commonly applied procedures only measure basal and maximal proton leak-linked oxygen consumption but not explicitly uncoupling protein 1 (UCP1)-dependent respiration.Meaningful oxygen consumption analyses require (1) Nebulisers the activation of UCP1, (2) control over intracellular free-fatty-acid levels, and (3) inhibition of ATP-consuming futile cycles.

For complete details on the use and execution of this protocol, please refer to Li et al.(2014, 2017, 2018) and Schweizer et al.(2018).