Influence Of FSHon Expression Of Several Related Genes Of Sertoli Cell Proliferation

Sertoli cells of testis are located in the seminiferous tubules, and it is the main component of spermatogenesis microenvironment. The proeess of SC proliferation is regulated by many hormones and factor, in all of which FSH is one of the most important hormones. Therefore, Study of FSH on the proliferation of regulatory mechanisms in vitro cultured sertoli cells of new born calves make a solid foundation for the number of Sertoli cells by artificial regulation, and also can improve male animal’s reproductive capacity.Sertoli Cells(SC), which can provide basis nutrition factors and high concentration of androgen for spermatogenic cells by synthesizing and secreting androgen-integrating protein, is thought to play a central role in spermatogenesis. Therefore, the research on SC would give us a better understanding on possible functions of SC and provide a promising application in male sterility and clinical transplantation.In this research, we used calves testis as basic material. SC were isolated by pancreatic enzyme and collagenase two-step method, purified by Percoll discontinue density gradient centrifugation, and identified by Feulgen staining, HE staining and Oil Red O staining. Different concentrations of FSH(0, 0.01, 0.02, 0.04, 0.08 IU/ml) were taken to treat sertoli cells in vitro culture, the proliferation of sertoli cells at six time points with five concentration were determined by CCK8,and then the expression of CDC25 gene was determined by Realtime-PCR.The two-step method could isolate and purify SC from calves testis, whose degree of purity was more than 95% after testing by Feulgen staining. SC which were cultured in vitro could survive to the 20 th generation and the rate of proliferation became down with time.FSH promoted the proliferation of sertoli cells and 0.04 IU/ml FSH concentration could significantly promoted the proliferation of in vitro cultured sertoli cells.FSH could significantly improved the gene expression in Signaling pathway of WNT/β-catenin, and could promote the proliferation of sertoli cells.FSH had no significant effect on expression the of CDC25 A, Within the range of certain concentration of FSH, FSH could significantly improved the expression of CDC25 B. CDC25 B might promoted the proliferation of sertoli cells, and decided the proliferation rate of sertoli cells.Within the range of certain concentration of FSH, FSH could significantly improved the expression of P53. P53 might controlled the proliferation; FSH could significantly decreased the expression of FAS and CDC25 C,and FSH could significantly improved the expression of FASL.