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http://dx.doi.org/10.5352/JLS.2014.24.12.1301

Composition of a Medium for Serum-free Culture of an Adipose-derived Stem Cell Line Established with a Simian Virus 40 T Antigen  

Kim, Gyu Bin (Department of Microbiology, College of Natural Sciences, Changwon National University)
Joo, Woo Hong (Department of Biology, College of Natural Sciences, Changwon National University)
Kim, Dong Wan (Department of Microbiology, College of Natural Sciences, Changwon National University)
Publication Information
Journal of Life Science / v.24, no.12, 2014 , pp. 1301-1307 More about this Journal
Abstract
Adipose-derived stem cells (ADSCs) are considered promising tools for tissue regeneration. However, ADSCs have very poor proliferation capacity. Therefore, fetal bovine serum (FBS) is generally added to the culture media of ADSCs. As FBS contains many uncharacterized components that may affect cellular functions, methods for serum-free cultures of ADSCs have been widely investigated. In this study, to develop an efficient method for a serum-free culture of ADSC-T, we used an ADSC line established by introducing the simian virus 40 (SV40) T gene into primary ADSCs. We then investigated the effect of amino acids, vitamins, and other components on the growth of ADSC-T. When the ADSC-T cells were plated with DMEM/F12 serum-free medium, the cells did not proliferate, and the mixture of amino acids, vitamins, and B27 supplement did not increase the growth of the cells. However, when the ADSC-T cells were provided with serum-free DMEM/F12 after they had been cultured with serum-supplemented DMEM for 24 h, the cells proliferated, and the vitamins and B27 supplement increased the cell growth. Stem-Pro serum-free medium also appeared to be useful as a suspension culture for the ADSC-T cells. The ADSC-T cells secreted large amounts of proteins of around 70 kDa. Insulin-like growth factor (IGF) and fibroblast growth factor basic (FGF basic) were secreted by ADSC-T in larger amounts in the serum-free culture than in the serum-supplemented culture.
Keywords
Adipose-derived stem cells; amino acids; cell culture; serum-free culture; vitamine;
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