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 µm, 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 µm, respectively), experimental 2 (15.6±3.5 and 157.6±41.9µm, respectively) groups were better than control 1 group (3.1±2.2 and 87.5±30.6µm, 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.