Materialandmethods分析.docVIP

Material and methods Cell Culture ADSC were isolated from adipose. Inguinal adipose tissue was excised from C57Bl/6 mice and digested at 37°C in Hanks buffer containing 2% BSA and 2 mg/ml collagenase (Worthington, Lakewood, NJ) for 60 min. After elimination of undigested fragments by filtration through a 40-?m cell strainer (BD Falcon, Bedford, MA), mature adipocytes were separated by centrifugation (280g, 10 minutes) and incubated in hemolysis buffer (160 mmol/L NH4Cl, 14 mmol/L NaHCO3, 0.1 mmol/L EDTA, pH 7.3) for 5 minutes. After centrifugation (280g, 10 minutes), ADSC were plated in DMEM