Effects of different concentrations of sucrose and fetal bovine serum on viability rate of lamb spermatogonial stem cells before and after cryopreservation

Spermatogonial stem cells (SSCs) have diverse applications in reproductive medicine and biotechnology. Cryopreservation is the well-known method for long-term storage of these cells. The aim of this study was to investigate the effect of different concentrations of fetal bovine serum (FBS) and Sucrose on the viability rate of spermatogonial cells.
Testicular cells of pre-pubertal lambs were separated in a two-step enzymatic isolation and purified by differential plating. Then, the cells were divided in 6 groups. Groups 1, 2 and 3 were treated with FBS (10%) and Sucrose (0.07, 0.14 and 0.21 molar concentration (M) ) and groups 4, 5 and 6 with FBS (20%) and Sucrose (0.07, 0.14 and 0.21 M), respectively. The viability rate of the cells was evaluated immediately after isolation, addition of cryoprotectant agents and thawing procedures. Identification of spermatogonial cells in the culture was performed using the immunocytochemistry staining against PGP9.5.
The results showed that cryoprotectant do not have any harmful effects on lamb´s SSCs. Moreover, viability rate of the cells in freezing media containing FBS (10%) is significantly higher than the media containing FBS (20%). Furthermore, increasing concentrations of Sucrose (0.07, 0.14 and 0.21 M) had no beneficial effect on the spermatogonial viability rate.
It was concluded that freezing media containing dimethyl sulfoxide (10%) and FBS (10%) and Sucrose (0.07 M) is appropriate for cryopreservation of lamb spermatogonial cells.