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CDC25A, TSSK3 and P53 expressionsin vitro in cultured sertoli cells after FSH treatment were studied in order to provide some data for further researches of spermatogenesis. Different concentrations of FSH (0, 0.01, 0.02, 0.04, and 0.08 IU? mL-1) were used to treat sertoli cells culturedin vitro. The expression of CDC25A, TSSK3 and P53 was determined by real-time-PCR at 6 h, 12 h and 24 h after FSH treatment of sertoli cells. The results showed that FSH had no significant effect on expression of CDC25A (p>0.05), could significantly improve the expression of TSSK3 and P53 (p<0.05), and had no significant effect on expression of CDC25A in sertoli cells, but it could significantly improve the expression of TSSK3. CDC25A was likely to play a role in other signaling pathways in sertoli cells. Within the range of certain concentration of FSH, TSSK3 in sertoli cells had the highest expression at about 24 h. TSSK3 protein produced in sertoli cells was likely to play an important role in substrate-level phosphorylationbe in meiosis and mitosis of spermatogenic cells. FSH could promote P53 expression and the highest expression was at about 12 h, and P53 might control the division of spermatogenic cells as well as sertoli cells.