Changes

*'''DMEM ''' (4.5 g/L glucose + L-glutamine – sodium pyruvate; Invitrogen catalog # 11965-092)*'''FBS ''' (we test several batches from several sources for differentiation and insulin stimulated glucose uptake then order large quantities of that particular lot. Currently we are using Sigma cat# F2442, lot# 072K8403. Aliquot and store in 50 mL tubes at -20)*'''NCS ''' (currently we are using Biowhittaker Cambrex cat# 14-416F, lot# 01106776; aliquot in 50 mL tubes and store at -20)*'''PSG ''' (100X Invitrogen Cat# 100378-016)*'''Insulin ''' (Sigma I-5523). Dissolve in 0.01N Hydrochloric acid as a 1 mg/ml stock. Aliquot into 1.5 mL tubes and store at -20*'''Dexamethasone ''' (Sigma D-1756). Dissolve in ethanol as a 2.5 mM stock. Aliquot into 1.5 tubes and store at -20.*'''MIX ''' (Sigma I-5879). Dissolve 2.78g/50mL KOH (0.98g/50mL). Aliquot into 1.5 mL tubes and store at -20*'''Fibroblast Media ''' (DMEM + 10% NCS + 1X PSG; Sterile filter into new bottle)*'''Adipocyte Media ''' (DMEM + 10 % FBS + 1X PSG; Sterile filter into new bottle)*'''Differentiation Media I ''' (Adipocyte Media + Insulin (1000X dilution), Dexamethasone (10 000X dilution) and MIX (500X dilution); Sterile filter into new bottle after additions*'''Differentiation Media II ''' (Adipocyte Media + Insulin (1000X dilution); Sterile filter into new bottle after adding insulin)