Expression profile of germ stem cell-specific genes in human spermatogonial stem cells after co culture with sertoli cells

Human spermatogonial stem cells (SSCs)، are the foundation of spermatogenesis. Because of low number and lack of significant marker in human SSCs، studying their characteristics، could provide better understanding about the biology of male fertility. This study was designed to examine the effects of in vitro co-culture with sertoli cells on SSC colonization and germ cells specific gene expression of human spermatogonial stem cells. Testicular cells were isolated from testis biopsies by using two step enzymatic digestion and differential plating. two culture system were designed: co-culture with patient Sertoli cells and culture of SSC without co-culture (as control group). The number and diameter of colonies were evaluated during 3 weeks of culture. The expression of alpha 6 integrin، beta1 integrin and PLZF، as germ stem cell specific markers، was assessed using quantitative RT-PCR. Statistical analysis was performed using one way ANOVA in SPSS vesion 16 software with 95% Confidence interval. Our results were showed that the number and diameter of colonies increased significantly in co-culture with sertoli cells (P<0. 05). The expression profile of genes in 2nd and 3rd weeks of culture revealed that there is significant higher expression of germ stem cell markers in our co-culture group versus control group. Based on the optimal effects of sertoli cells on spermatogonial stem cells، co culture of the human SSCs with the feeder layer sertoli may be used as a suitable method for the enrichment of human spermatogonial stem cells.