Influence Factors of in vitro Culture of Goat Spermatogonial Stem Cells

This study aims to determine the condition factors influencing the in vitro proliferation culture of spermatogonial stem cells (SSCs) through isolation and culture of SSCs. With mouse embryonic fibroblast (MEF) cells, SIM-6-thiogunanie-oualiain (STO) cells and Hela cells used as the feeder layers, we compared the numbers of cell clones treated with mitomycin C as well as the proliferation conditions with and without the treatment with trypsin. In the culture solution, we respectively added serum and vitamin C (of different concentrations, VC) to culture goat SSCs; furthermore, we tested their pro-proliferation effect on goat SSCs and its mechanism. We determined that one-month-old goats were the most suitable for the in vitro culture; 1.5 h, 2.5 h and 2.5 h were respectively the most suitable treatment times in the feeder layers of MEF cells, STO cells and Hela cells; 2.5% was the optimal serum concentration; VC (30 μg/mL) had a promoting effect on the in vitro culture of goat SSCs. We conclude that the age of goats, different feeding layers and their preparation conditions, serum concentrations and VC concentrations are the condition factors influencing the in vitro culture and proliferation of SSCs.