Effect of cortisol on caspases in the co-cultured C2C12 and 3 T3-L1 cells.

Abstract

The present study was carried out to understand the effect of cortisol on caspase expression in the C2C12 and 3T3-L1 cells under co-culture system. Cells were co-cultured by using transwell inserts with a 0.4-μm porous membrane to separate C2C12 and 3T3-L1 preadipocyte cells. Each cell type was grown independently on the transwell plates. Following cell differentiation, inserts containing 3T3-L1 cells were transferred to C2C12 plates and inserts containing C2C12 cells were transferred to 3T3-L1 plates. A total of 10μg/μl of cortisol was added to the medium. Following treatment of cortisol for 3days, the cells in the lower well were harvested for analysis. Caspases such as caspase 3, caspase 7, and caspase 9 were selected for the analysis. qRT-PCR results indicated the significant increase in the mRNA expression of caspase 3, caspase 7, and caspase 9. Caspase 3, 7, and 9 activities were also increased in the mono- and co-cultured C2C12 and 3T3-L1 cells. In addition, confocal microscopical investigation indicated that cortisol increases caspase expressions in the mono- and co-cultured C2C12 and 3T3-L1 cells. Taking all these together, we concluded that the co-culture system reflects the exact effect of cortisol on caspase expression, which is quite distinct from one dimensional mono-cultured experiments.