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Effect of Cortisol on Caspases in the Co-cultured C2C12 and 3 T3-L1 Cells

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Abstract

The present study was carried out to understand the effect of cortisol on caspase expression in the C2C12 and 3 T3-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 3 T3-L1 preadipocyte cells. Each cell type was grown independently on the transwell plates. Following cell differentiation, inserts containing 3 T3-L1 cells were transferred to C2C12 plates and inserts containing C2C12 cells were transferred to 3 T3-L1 plates. A total of 10 μg/μl of cortisol was added to the medium. Following treatment of cortisol for 3 days, 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 3 T3-L1 cells. In addition, confocal microscopical investigation indicated that cortisol increases caspase expressions in the mono- and co-cultured C2C12 and 3 T3-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.

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Acknowledgments

This article work was supported by research funds of Catholic University of Daegu in 2014.

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  1. Department of Food Science and Nutrition, Catholic University of Daegu, Daegu, South Korea

    Pandurangan Muthuraman

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  1. Pandurangan Muthuraman
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Correspondence to Pandurangan Muthuraman.

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Muthuraman, P. Effect of Cortisol on Caspases in the Co-cultured C2C12 and 3 T3-L1 Cells. Appl Biochem Biotechnol 173, 980–988 (2014). https://doi.org/10.1007/s12010-014-0909-z

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  • DOI: https://doi.org/10.1007/s12010-014-0909-z

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