摘要
本研究采用腺病毒感染、慢病毒感染、脂质体转染和电穿孔转化方法将含有绿色荧光蛋白(GFP)的质粒转入经过差异贴壁法初步分离纯化的小鼠精原干细胞(SSCs)中,转染48 h后通过流式细胞仪检测GFP阳性细胞比例比较4种方法在体外转染精原干细胞的效率.结果显示,脂质体转染效率最高仅为8.64%,不能满足对精原干细胞进一步实验的要求;电穿孔法效率最高达到25.27%,但转化后细胞大量死亡;腺病毒转染细胞的效率达到了32.4%;慢病毒转染效率最高,达到74.25%.因此,慢病毒转染法是体外转染小鼠精原干细胞的有效方法.
In this study, we compared different methods using adenovirus, lentivirus, liposomes and electroporation for efficient transduction of purified spermatogonial stem cells （SSCs）. The expression of green fluorescence protein （GFP） were assayed by flow cytometry after cell treatments for 48 hours. The highest efficiency of liposome transfection was low as 8.64%. The electroporation reached 25.27% with significant of cell death. The maximum efficiency of adenoviral infection was observed to be 32.4%. The lentivirus infection gave the best efficiency of 74.25%. We concluded that the most suitable method to transduce SSCs in vitro was to use lentivirus as the gene transfer vectors.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2014年第1期93-100,共8页
Chinese Journal of Biochemistry and Molecular Biology
基金
国家重点基础研究发展计划项目(973计划
No.2011CBA01002)~~
关键词
精原干细胞(SSCs)
转染效率
不同转染方法比较
permatogonial stem cells（SSCs）
transfection efficiency
different transfection methods
