Radiotherapy and chemotherapy could affect spermatogenesis process, so it is important to preserve fertility of patients who suffer from cancer disease. Cryopreservation of SSCs is considered as a helpful procedure for preserving male fertility in cancer patients. It seems that antioxidants can decrease ROS formation and reduce side-effects of cryopreservation on cells. In this study, we added catalase and alpha-tocopherol to the basic freezing medium in order to evaluate the effects of these two antioxidants on SSCs.
Materials and methods
SSCs were isolated from testes of 3 to 6 days old mice using enzymatic digestion. The purity of isolated cells was evaluated by flow cytometry. Catalase (20 and 40 μl/ml) or alpha-tocopherol (100 and 200 μl/ml) was added to the basic freezing medium. The cell viability, ROS formation and expression of Bax and Bcl2 were evaluated in these conditions.
The survival rate of the freezed cells in the presence of catalase or alpha-tocopherol was significantly more than control group (P≤0.05). In addition, the Bax expression level in the catalase and alpha-tocopherol groups was significantly (P≤0.05) lower than the control group and a significant rise of Bcl2 expression was detected in the catalase and alphatocopherol groups (P≤0.05). However, the ROS production in catalase and alpha-tocopherol was significantly (P≤0.05) lower compared with the control group.
This cryopreservation method could help to increase the number of SSCs and improve the quality and viability of these cells after cryopreservation