Nutrition medium

Human primary preadipocytes prepared from liposuction material were obtained from PromoCell and differentiated as previously described [10 ]. In brief, cells were expanded in growth medium (PromoCell) and differentiation (day 0) was initiated by switching for three days to differentiation medium (PromoCell). Thereafter, the cells were cultured in nutrition medium (PromoCell) containing glibenclamide, glimepiride, rosiglitazone or SR1664 (all Sigma–Aldrich) dissolved in DMSO until day 21 if not otherwise stated. 3T3-L1 preadipocytes were obtained from the American Type Culture Collection (ATCC) and were cultured in Dulbecco's modified Eagle's medium (DMEM, Invitrogen) and 10% fetal bovine serum. Differentiation was induced as previously reported [29 (link)]. glibenclamide, glimepiride, rosiglitazone or SR1664 were added to culture media from induction of differentiation on (day 0) until day 7 if not otherwise stated. Human (Sigma–Aldrich) or murine (Miltenyi Biotec) tumor necrosis factor α (TNFα) were dissolved in water and added to culture media as indicated.

3T3-L1, HeLa cells were obtained from ATCC, cultured in DMEM with 10% FBS and 1% penicillin/streptomycin/L-glutamine (Biological Industries). 3T3-L1 cells were differentiated (12–14 days) into mature adipocytes^{79 (link)}. LiSa-2 cells were a gift from Peter Moeller (University of Ulm, Germany), were cultured in DMEM/F12 (1:1) with HEPES, 10% FBS and 1% penicillin/streptomycin (Biological Industries) and differentiated (7 days) into adipocyte-like cells⁸⁰ . H1299 cells with homozygous partial deletion of p53 and deficient in p53 protein expression were obtained from ATCC and maintained in RPMI 1640 medium supplemented with 10% FBS and 1% penicillin/streptomycin (Biological Industries). Primary human white subcutaneous pre-adipocytes (HWP; PromoCell, female donor) (Cat# C12730; Lot # 419Z023) were cultured in pre-adipocyte growth medium (PromoCell). At 80–90% confluence, differentiation was induced by differentiation medium (PromoCell) for 3 days, followed by culturing in a nutrition medium (PromoCell) that was renewed every 2 days for 6–8 days.