Culture and Identification of Testis Sertoli Cells in Calf in vitro

In order to explore the cultural characteristics of sertoli cells in vitro,calf testis were used in the present study.The method of collagenase and typin was sequentially performed to separate sertoli cells,and then the differential adhesion method was used to purify sertoli cells.The sertoli cells obtained were cultured in vitro and identified.The results showed that the sertoli cells could be passaged to the 20th generation,and with the increase of the number of passages,cell proliferation ratio was getting slower and slower;DMEM/F-12 medium containing 2% fetal bovine serum could be used to culture sertoli cells in vitro;Immunohistochemical staining result indicated that sertoli cells expressed vimentin.The marker gene SCF and GDNF could be detected by RT-PCR analysis in sertoli cells.The 15th generation of sertoli cells contained normal diploid karyotype.This study not only identified calf sertoli cells effectively,but also laid foundation for the probe into the physiological function of sertoli cells cultured in vitro.