Culturing and Differentiating Murine Brown Adipocytes

The murine non-differentiated brown adipocyte cell line [18 (link)] (kindly provided by Dr A. Whittle, Institute of Metabolic Science, University of Cambridge, UK) was cultured in Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 25 mM glucose, 10% fetal calf serum (FCS), 1% glutamine, 1% penicillin and 1% streptomycin in a humidified atmosphere at 37°C and 5% CO₂. Differentiation of the brown adipocyte cell line into mature brown adipocytes was conducted as described [18 (link),19 (link)]. The density of non-differentiated brown adipocytes depending on the further experimentation was determined by the Cellometer Auto T4 counter (Nexcelom Bioscience).