نیلوفر خرمی

The aim of this research was to assess the effect of melatonin on the diameter of colonies, apoptosis status and apoptosis related genes expression in sheep’s spermatogonial stem cells (SSCs). 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), melatonin (1 nmol) and melatonin+H2O2 (30 µM H2O2 along with 1 nmol melatonin), respectively. The cells were cultured for 3 weeks and the colonies’ diameter were evaluated on the 5th, 14th and 21th days of culture. At the end of culturing period, apoptosis status and apoptosis related genes expression (BAX and BCL2) were evaluated. On the 5th and 14th days of culture, the diameter of colonies were higher (P≤0.05) in the control and melatonin groups compared to the other groups. On the 21th day of culture, the highest and the least (P≤0.05) diameter of colonies were observed in melatonin and H2O2 groups, respectively. The least and the highest (P≤0.05) rate of apoptotic SSCs, BAX expression and BAX/BCL2 ratio were observed in melatonin and H2O2 groups, respectively. Moreover, the highest (P≤0.05) expression of BCL2 gene was found in melatonin group. In conclusion, using melatonin in culture medium could be an effective way to improve the quality and decrease apoptosis status in sheep’s SSCs.

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.

This study was conducted to evaluate the effect of IGF-I on the number and area of colonies, viability 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 control group did not receive IGF-I and the other groups received 0.1, 1, 5 and 10 μg/ml IGF-I, respectively. The cells were cultured for 2 weeks and the colonizing abilities were evaluated on the 5th and 14th days of culture. At the end of culturing period, apoptosis status and apoptosis related genes expression (BAX and BCL2) were evaluated. The results showed using 5 and 10 μg/ml IGF-I improved colonizing abilities on the 5th and 14th days compared to the other groups (P≤0.05). The highest viability and lowest apoptosis status was observed in groups received 5 and 10 μg/ml IGF-I (P≤0.05). The highest BCL2 expression and lowest BAX expression was observed in groups received 5 and 10 μg/ml IGF-I (P≤0.05). Therefore, using suitable dose of IGF-I in culture medium could be an effective method to improve the quality and viability of sheep’s SSCs during studying on stem cells.