ABSTRACT

Culturing cells on bio-gels are believed to provide a more in vivo-like extracellular matrix. 3T3-L1 cells cultured on Matrigel® significantly alteregd their proliferation and differentiation as compared to growth on tissue culture-coated polystyrene surfaces. Growth on a 250-μm thick layer of Matrigel® facilitated the formation of cellular aggregates of 3T3-L1 cells. Differentiation of 3T3-L1 cells cultured on Matrigel® demonstrated increased levels of mRNA levels for key adipogenic transcription factors (PPARγ, C/EBPα, SREBP1), lipogenic markers (FAS, FABP4, LPL, PLIN1) and markers of adipocyte maturity (LEP), compared to cells cultured directly on a polystyrene tissue culture surface. The gene expression of extracellular matrix proteins (FN1, COL1A1, COL4A1, COL6, LAM) was decreased in 3T3-L1 cells cultured on Matrigel®. Furthermore, growth on Matrigel® increased lipid accumulation in 3T3-L1 cells in the presence and absence of rosiglitazone, a thiazolidinedione routinely used to optimize differentiation in these cells. These changes in adipocyte gene expression and lipid accumulation patterns may be a result of the increased cell–cell and cell–ECM interactions occurring on the Matrigel®, a scenario that is more reflective of an in vivo model. Taken together, our data advance the understanding of the value of culturing 3T3-L1 cells on Matrigel®.

摘要

人们认为在生物凝胶上培养细胞可以提供更像体内的细胞外基质。与在组织培养物包被的聚苯乙烯表面上生长相比，在 Matrigel® 上培养的 3T3-L1 细胞显着改变其增殖和分化。在 250 μm 厚的 Matrigel® 层上的生长促进了 3T3-L1 细胞的细胞聚集体的形成。在 Matrigel® 上培养的 3T3-L1 细胞的分化表明，关键脂肪生成转录因子（PPARγ、C/EBPα、SREBP1）、脂肪生成标记物（FAS、FABP4、LPL、PLIN1 ）和脂肪细胞成熟标记物（ LEP）的 mRNA 水平增加，与直接在聚苯乙烯组织培养表面上培养的细胞相比。在 Matrigel® 上培养的 3T3-L1 细胞中，细胞外基质蛋白（FN1、COL1A1、COL4A1、COL6、LAM）的基因表达降低。此外，无论是否存在罗格列酮（一种常规用于优化这些细胞分化的噻唑烷二酮），Matrigel® 上的生长都会增加 3T3-L1 细胞中的脂质积累。脂肪细胞基因表达和脂质积累模式的这些变化可能是 Matrigel® 上细胞与细胞以及细胞与 ECM 相互作用增加的结果，这种情况更能反映体内模型。总而言之，我们的数据加深了对在 Matrigel® 上培养 3T3-L1 细胞价值的理解。