Lequeux C, Auxenfans C, Mojallal A, Sergent M, Damour O
Laboratoire des Substituts Cutanés, Hôpital Edouard Herriot, Lyon, France.
Biomed Mater Eng. 2009;19(4-5):283-91. doi: 10.3233/BME-2009-0593.
Our objective was to optimize a medium for preadipocyte differentiation into adipocytes.
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.
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.
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 microM rosiglitazone+0.15 UI/ml insulin+antibiotics.
我们的目标是优化一种用于前脂肪细胞分化为脂肪细胞的培养基。
分化培养基包含固定成分以及7种可变成分。为开展本研究，设计了不同实验，并分4个阶段进行。前两个阶段测试了血清、地塞米松、氢化可的松和一种环磷酸腺苷激活剂的影响。在第三阶段，添加了两个新变量：罗格列酮和胰岛素。在最后阶段，对第三阶段选择的培养基进行验证。分化选择标准包括成熟脂肪细胞数量和脂联素分泌。
我们已表明每个变量都是不可或缺的，并且一些变量之间存在正相互作用。未发现负相互作用，并且有可能优化每个变量的浓度。
我们选择了以下培养基，其可提供最佳的脂肪细胞大小和脂联素分泌：DMEM/HAMF12 + 10%胎牛克隆血清（FCS）+ 2 nM三碘甲状腺原氨酸+ 10 nM氢化可的松+ 0.5 mM异丁基甲基黄嘌呤（IBMX）+ 500 nM地塞米松+ 1 microM罗格列酮+ 0.15 UI/ml胰岛素+抗生素。
