Culture and differentiation of preadipocytes

LW Li Weng
WT Wen-Shuai Tang
XW Xu Wang
YG Yingyun Gong
CL Changqin Liu
NH Ni-Na Hong
YT Ying Tao
KL Kuang-Zheng Li
SL Shu-Ning Liu
WJ Wanzi Jiang
YL Ying Li
KY Ke Yao
LC Li Chen
HH He Huang
YZ Yu-Zheng Zhao
ZH Ze-Ping Hu
YL Youli Lu
HY Haobin Ye
XD Xingrong Du
HZ Hongwen Zhou
PL Peng Li
TZ Tong-Jin Zhao
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3T3-L1 preadipocytes and SVFs were cultured in Medium A at 37oC in an atmosphere of 8.8% CO2 and maintained in less than 50% confluence. Cells were subjected to a standard cocktail hormone as previously described49,51. After day 8, mature adipocytes were cultured in Medium B (Dulbecco’s modified Eagle’s medium (DMEM) with high glucose (4.5 g/L), 10% fetal bovine serum, 100 U/ml penicillin, and 100 mg/ml streptomycin), and fresh medium was changed every two days. For inducible deletion of Med20, SVFs-derived adipocytes (Rosa-CreERT2;Med20f/f), on day 6 of differentiation, were treated with 4-OHT (10 μM) for 4 days. On day 14, cells were harvested for imaging under bright field. The cultured medium was also collected for measurement of released LDH (Beyotime, #C0016).

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