In-vitro propagation of pre-pubertal bovine Spermatogonial Stem Cell in prepation for transplantation

‌Backgroud: We investigated the expression of stem cell markers in in -vitro cultivated speramatogonial stem cells (SSCs) with the intention of developing a useful system to produce progeny of SSCs. Materials and Mehtods: Testes from 3, 5 and 7 months old calves were utilized to examine isolation, purity and in –vitro propogation of SSCs. Cells were isolated and purified with two-stemp enzymatic digestion combined with centrifugal separation on discontinuous Percoll density gradient layers. Findings: Cell propagation and SSCs marker expression were determined at 5, 10 and 15 days post-culture. Immunostaining in conjunction with transcrioption based marker expression of cultured cells showed that stem cell markers (DBA, UCHL1, Oct4 and Sox2) were expressed in SSCs. Conclusion: The results demonstrated marker expression of bull spermatogonial stem cells and showed that prior to transplantation in-vitro culturing of bull SSCs is implementable.