Adipocyte differentiation is affected by media height above the cell layer.

3T3-L1 cells have been widely used as a model for adipogenesis. However, despite its popularity, differentiation of this cell line has been reported to be inconsistent with low efficiency. To investigate the effect of media height during adipocyte differentiation on lipid accumulation and adipokine secretion in mature adipocytes. Three cell lines (3T3-L1, OP9, and ChubS7) were used to test the influence of media volume on adipogenesis. Total lipid content and lipid droplet size and number were quantified. Adipocyte related gene expressions were quantified during the course of differentiation. Secretion of leptin and adiponectin from mature adipocytes were measured using Enzyme-Linked Immunosorbent Assays. The influence of oxygen partial pressure on adipogenesis was investigated using three oxygen percentages: 5%, 21%, and 30%. Insulin sensitivity was measured by insulin inhibition of isoproterenol-induced lipolysis and phosphorylation of Insulin Receptor Substrate-1 (IRS-1). A lower media height during adipogenesis increased total lipid accumulation, NEFA release and leptin and adiponectin secretion in mature adipocytes. Insulin sensitivity was not affected by media height during differentiation. Media height during adipogenesis was inversely correlated with lipid content in mature adipocytes. To achieve a high lipid content and greater adipokine secretion, it is best to use a low media volume during differentiation.

3T3 - L1细胞已被广泛用作脂肪生成的模型。然而，尽管它很常用，但据报道该细胞系的分化效率低下且不稳定。 为了研究脂肪细胞分化过程中培养基高度对成熟脂肪细胞中脂质积累和脂肪因子分泌的影响。 使用了三种细胞系（3T3 - L1、OP9和ChubS7）来测试培养基体积对脂肪生成的影响。对总脂质含量以及脂滴大小和数量进行了定量。在分化过程中对脂肪细胞相关基因的表达进行了定量。使用酶联免疫吸附测定法测量了成熟脂肪细胞中瘦素和脂联素的分泌。使用三种氧气百分比（5%、21%和30%）研究了氧分压对脂肪生成的影响。通过胰岛素对异丙肾上腺素诱导的脂解的抑制以及胰岛素受体底物 - 1（IRS - 1）的磷酸化来测量胰岛素敏感性。 在脂肪生成过程中较低的培养基高度增加了成熟脂肪细胞中的总脂质积累、非酯化脂肪酸（NEFA）释放以及瘦素和脂联素的分泌。分化过程中培养基高度不影响胰岛素敏感性。 脂肪生成过程中的培养基高度与成熟脂肪细胞中的脂质含量呈负相关。为了实现高脂质含量和更多的脂肪因子分泌，在分化过程中最好使用低体积的培养基。