The Optimization of Isolation and Culturing of Spermatogonial Stem Cells in Mouse

Testicular cells of 5-7 days postpartum Kunming mouse were collected by two-step enzymatic digestion method. Effects of two different purification methods known as different adherence selection and percoll discontinuous density gradients were compared, whereas the effects of different growth factors on spermatogonial stem cells (SSCs) proliferation were investigated as well by MTT assay. The results showed that there was significant difference in the numbers of total testes cells acquired between 5 days postpartum pups and 7 days postpartum pups(P＜0.05), yet the numbers of SSCs had no significant difference between the two groups(P＞0.05).The purification method of different adherence selections attained higher SSCs numbers and purification ratio than Percoll method, with the ratio of 85.06%.By means of MTT assay, the results showed that LIF factor could promote SSCs proliferation, and the 20 ng/mL GDNF+20 ng/mL bFGF+10³ U/mL LIF was the best supplementation combination in Kunming mouse SSCs in vitro.