Colony formation ability of frozen thawed spermatogonial stem cell from adult mouse

Background: The basis of spermatogenesis is the spermatogonial stem cells (SSCs). The concentration of SSCs is very small. However, a system that supports the proliferation and maintenance of SSCs in vitro could be used to preserve and expand SSCs numbers as well as increase success in transplantation. It is a new avenue to restore spermatogenesis in azoospermia subjects. Objective: Proliferation and enhancement of frozen-thawed SSCs numbers during in vitro culture. Materials and Methods: Both Sertoli and spermatogonial cells were isolated from adult mouse testes. Frozen-thawed spermatogonial cells were cultured in two groups: simple culture (Experimental 1) and co culture with Sertoli cells (Experimental 2). Also, Fresh cells were considered as control groups: simple culture (control1) and co culture with Sertoli cells (control 2).Assay of the spermatogonial-cell-derived colonies was carried out at the end of each week. Results: Results indicated that the viability rate of the frozen cells after thawing (68.4±10.2%) was influenced by cryopreservation procedure significantly (p ?0.001). In addition, the number of the colonies and their diameters in the co-culture system with fresh cells (25.1±5.2 and 205.8±50 ?m, respectively) were more than other groups and the differences were significant (p < 0.001). Number of the colonies and their diameters in experimental 1(9.5±4.3 and 124±35.9 ?m, respectively), experimental 2 (15.6±3.5 and 157.6±41.9?m, respectively) groups were better than control 1 group (3.1±2.2 and 87.5±30.6?m, respectively) and the differences were significant (p < 0.001). Conclusion: We demonstrated that co-culture system with Sertoli cells can increase in vitro colony formation of adult fresh and frozen-thawed spermatogonial cells in mouse.