Effect of testosterone on the differentiation control of stromal vascular cells isolated from longissimus muscle of Hanwoo beef cattle

Testosterone, as an influential factor in marbling score, requires strict management for uniform development of adipocytes in-between muscle bundles. Present study investigated effect of castration timing and testosterone levels on adipocyte development using SVCs. Isolated SVCs exhibited classical MSC markers, CD31(-), CD34(-), CD45(-), CD90(+), and CD105(+). ELISA analysis indicated that serum testosterone concentration was highest in non-castrated calves while no significant difference was observed between female, early and late castrated calves. CCK-8 assay showed that concentration of testosterone had no effect on cell proliferation. However, the real-time PCR demonstrated that 20 ng/ml of testosterone suppressed expression of preadipocyte markers, pref-1 and zfp423, but encouraged expression of myoblast markers, myf5 and myoD, via the AR. Consequently, expression of adipogenic markers C/EBP alpha and PPAR gamma, as well as accumulation of triglyceride, were decreased in 20 ng/ml testosterone treatment under adipogenic conditions. These findings suggest that by castrating calves before level of testosterone increases, may improve marbling development in the Hanwoo beef industry.