Optimization of a culture medium for the differentiation of preadipocytes into adipocytes in a monolayer

Our objective was to optimize a medium for preadipocyte differentiation into adipocytes. Methods: The differentiation medium contains fixed components as well as 7 variable ones. To perform this study, different experiments were designed and the study was carried out in 4 stages. The first two stages tested the influence of serum, dexamethasone, hydrocortisone and an cAMP activator. In the third stage, two new variables were added: rosiglitazone and insulin. In the final stage, the medium selected in stage 3 was validated. The differentiation selection criteria consisted of the number of mature adipocytes and adiponectin secretion. Results: We have shown that each variable was indispensable and that positive interactions occurred between some variables. No negative interactions were found and it was possible to optimize the concentration of each variable. Conclusions: We selected the following medium, which provides optimal adipocyte size and adiponectin secretion: DMEM/HAMF12 + 10% Foetal Clone Serum (FCS) + 2 nM triiodothyronine + 10 nM hydrocortisone + 0.5 mM IsoButyl Methyl Xanthine (IBMX) + 500 nM dexamethasone + 1 mu M rosiglitazone + 0.15 UI/ml insulin + antibiotics.