摘要
目的比较胎牛血清和驴自体血清对体外培养驴卵泡颗粒细胞增殖和凋亡的影响,从而提高驴卵泡颗粒细胞的培养效率。方法本试验将第三代贴壁的驴卵泡颗粒细胞分别用含10%胎牛血清与含5%、10%、15%驴自体血清的培养液进行对比培养。MTT法与划痕实验测定细胞的增殖和迁移,流式细胞技术检测其凋亡,酶标仪检测细胞SOD活力,RT-qPCR分析凋亡与自噬相关基因BAX、BCL-2、ATG7表达水平。结果各组间细胞增殖无显著差异,但驴自体血清各组细胞的迁移能力均优于胎牛血清组;驴自体血清各组细胞的凋亡率极显著低于胎牛血清组(P<0.01);10%自体血清组,SOD活力最高,基因BAX、ATG7表达极显著低于其他各组(P<0.01),而基因BCL-2表达则相反。结论含驴自体血清的培养液对驴卵泡颗粒细胞的增殖、迁移及活力优于含胎牛血清的培养液。
Objective The aim of study is to compare the effects of fetal bovine serum and donkey autologous serum on the proliferation and apoptosis of donkey follicular granulosa cells cultured in vitro,so as to improve the cultural efficiency of donkey follicular granulosa cell.Methods The third generation adherent donkey follicular granulosa cells were cultured in the medium containing 10%fetal bovine serum and 5%,10%and 15%donkey autologous serum respectively.MTT assay and scratch test were used to detect cell proliferation and migration.Flow cytometry was used to detect apoptosis.Microplate reader was used to detect SOD activity of cells.RT-qPCR was used to analyze the expression level of apoptosis and autophagy related genes BAX,BCL-2 and ATG7.Results There was no significant difference in cell proliferation among groups.The migration ability of donkey autologous serum groups were all better than that of fetal bovine serum group,which was contrary in the apoptosis rate(P<0.01).The SOD activity was the highest,and the expression of Bax and Atg7 in 10%autologous serum group was significantly lower than that of other groups(P<0.01),while the expression of Bcl-2 was on the contrary.Conclusion The culture medium containing donkey autologous serum had better proliferation,migration and vitality of donkey follicular granulosa cells than the culture medium containing fetal bovine serum.
作者
郭旭川
王艳萍
张强
王璐
曾维斌
GUO Xuchuan;WANG Yanping;ZHANG Qiang;WANG Lu;ZENG Weibin(College of Animal Science and Technology,Shihezi University,Shihezi,Xinjiang 832000,China)
出处
《石河子大学学报（自然科学版）》
北大核心
2021年第4期440-445,共6页
Journal of Shihezi University(Natural Science)
基金
国家自然科学基金项目(31360530,31760643)
石河子大学创新发展专项(CXFZ202007)。
关键词
驴
颗粒细胞
血清
增殖
凋亡
donkey
granular cells
serum
proliferation
apoptosis
