Effect of Calligonum extract on the colonizing ability and ROS production in sheep’s spermatogonial stem cells

This study was aimed to investigate the effect of Calligonum extract on the number of colonies, ROS production and apoptosis related genes expression in sheep’s spermatogonial stem cells (SSCs). In this study, SSCs at the basal membrane of seminiferous tubules were isolated from testes of Afshari sheep using enzymatic digestion steps. The samples assigned into four groups. The control group received basic medium and the other groups contained H2O2 (30 µM), Calligonum (10 µg/ml) and Calligonum+H2O2 (30 µM H2O2 along with 10 µg/ml Calligonum), respectively. The cells were cultured for 3 weeks and the colonizing abilities were evaluated on the 5th, 14th and 21th days of culture. At the end of culturing period, ROS production and apoptosis related genes expression (BAX and BCL2) were evaluated. On the 5th and 14th days of culture, the number of colonies were higher (P≤0.05) in the control and Calligonum groups compared to the other groups. On the 14th day of culture, the highest and the least (P≤0.05) number of colonies were observed in Calligonum and H2O2 groups, respectively. The least and the highest (P≤0.05) rate of ROS contained SSCs, BAX expression and BAX/BCL2 ratio were observed in Calligonum and H2O2 groups, respectively. Moreover, the highest (P≤0.05) expression of BCL2 gene was found in Calligonum group. In conclusion, using Calligonum extract in culture medium could be a helpful strategy to improve the quality and decrease apoptosis status in sheep’s SSCs during research studies.