Sertoli cells from mouse testis were isolated and cultured to study the effect of Sertoli cells feeders on mouse spermatogonial stem cells in vitro.Sertoli cells and spermatogonial stem cells were isolated from 3~7 d ICR mouse and subjected to sequential enzymatic treatment.Sertoli cells were treated with Tris-HCl to remove spermatogenic cells and high purity sertoli cells were obtained.Sertoli cells treated with mitomycin C were used as feeder cells,to study the effect on mouse spermatogonial stem cells.After a week of culture,compared with control,there were more number of spermatogonial stem cells remained.More than 90% of the Sertoli cells were isolated and cultured.Sertoli cells can promote proliferation of spermatogonial stem cells obviously.More Sertoli cells were obtained through this experiment method with high purification.Sertoli cells can be used as feeders not only to promote survival but also renewal of spermatogonial stem cells.