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

The basis of spermatogenesis is the spermatogonial stem cells (SSCs).The concentration of SSCs is very small. However, a system that supports theproliferation and maintenance of SSCs in vitro could be used to preserve and expandSSCs numbers as well as increase success in transplantation. It is a new avenue torestore spermatogenesis in azoospermia subjects.

Proliferation and enhancement of frozen-thawed SSCs numbers during in vitroculture.

Both Sertoli and spermatogonial cells were isolated fromadult mouse testes. Frozen-thawed spermatogonial cells were cultured in two groups: simpleculture (Experimental 1) and co culture with Sertoli cells (Experimental 2). Also, Fresh cellswere 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 endof each week.

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). Inaddition, the number of the colonies and their diameters in the co-culture system withfresh cells (25.1±5.2 and 205.8±50 μm, respectively) were more than other groups andthe differences were significant (p<0.001). Number of the colonies and their diametersin experimental 1(9.5±4.3 and 124±35.9 μm, respectively), experimental 2 (15.6±3.5and 157.6±41.9μm, respectively) groups were better than control 1 group (3.1±2.2 and87.5±30.6μm, respectively) and the differences were significant (p<0.001).

We demonstrated that co-culture system with Sertoli cells can increase invitro colony formation of adult fresh and frozen-thawed spermatogonial cells in mouse.