小鼠精原干细胞系自我更新和分化能力的体外观察  

In vitro observation of self-renewal and differentiation capacity of mouse spermatogonial stem cell line

作　　者：薛江阳 高亭亭[1] 邵彬彬 郑皓宇[1] 郑波[1] 黄晓燕[1] 

机构地区：[1]南京医科大学组织胚胎学系,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2016年第2期155-159,共5页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家重点基础研究发展计划(973计划)(2011CB944301)

摘　　要：目的 :体外观察小鼠精原干细胞(spermatogonial stem cells,SSCs)的自我更新和分化能力。方法 :体外构建稳定的小鼠SSCs系,利用Ed U细胞增殖分析试剂盒检测小鼠SSCs体外自我更新的能力;利用活细胞工作站进一步观察小鼠精原干细胞体外增殖现象;运用TUNEL法检测SSCs凋亡现象;通过维甲酸(retinoic acid,RA)诱导小鼠SSCs,观察其体外分化的能力。结果:Ed U孵育小鼠SSCs 2 h后,流式检测细胞增殖率平均为40.75%;活细胞工作站下可见正在分裂的干细胞;TUNEL检测显示干细胞中凋亡信号很少;RA诱导后,标记SSCs分化的基因(C-kit、Scp3和Stra8)表达量明显升高(P<0.05)。结论:稳定构建的小鼠SSCs系与体内的SSCs类似,具有较强的自我更新以及细胞分化能力,为下一步开展精原细胞相关基因和蛋白功能研究提供了良好的技术平台。Objective：To observe the ability of self-renewal and differentiation of mouse spermatogonial stem cells（SSCs）in vitro.Methods：Mouse spermatogonial stem cell line was established stably. The capacity of SSCs self-renewal in vitro was detected via EdU Proliferation Kit; Cell proliferation was observed by Live cell Imaging System. Cell apoptosis was tested by TUNEL method. The capacity of SSCs differentiation was observed by retinoic acid（RA） induction. Results：After incubation with EdU for 2 hours,the proliferation rate was analyzed as an average of 40.75% by EdU Flow Cytometry Assay Kits. The dividing stem cells were clearly catched via short-term live cell imaging. The signal of apoptosis of SSCs was nearly undetected by TUNEL methods. The expression of genes（C-kit,Scp3 and Stra8）marking differentiation was increased after RA induction（P〈0.05,n=3）. Conclusion：Similar as SSCs in vivo,SSC line in vitro also has the capacity of self-renewal and differentiation. It provides us a good platform to carry out the function of the related genes and proteins during mouse spermatogenesis.

关 键 词：精原干细胞 自我更新 分化 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]