猪耳成纤维细胞的体外培养及EGFP基因的转染  

In vitro culture of pig ear skin fibroblasts and transfection of EGFP gene

作　　者：刘伟[1] 吴华莉[1] 张德福[1] 

机构地区：[1]上海市农业科学院畜牧兽医研究所

出　　处：《上海农业学报》2007年第4期10-13,共4页Acta Agriculturae Shanghai

基　　金：上海市科技兴农重点攻关项目[沪农科攻字(2005)第14-1号];上海市科学技术委员会项目(01490900504DZ05611);上海市自然科学基金项目(98ZC14025)

摘　　要：优化脂质体Liperfectamin 2000转染EGFP（加强型绿色荧光蛋白）至猪耳成纤维细胞的参数，如DNA和脂质体的用量、细胞汇合度、细胞暴露于DNA和脂质体复合物的时间。试验显示：24孔板内1．0μg DNA和2．4μL脂质体的用量转染效率最高，DNA和脂质体过量会对细胞产生毒性，影响转染效率；细胞汇合85％～90％才能得到较高的转染效率；细胞暴露4h转染效率最高，时间过长反而使转染效率下降。The transfection of EGFP （enhanced green fluorescent protein） by liposome Liperfectamin 2000 into pig ear skin fibroblasts was investigated in such parameters as the doses of DNA and liposome, the cell confluence and the time of exposing the cells to the DNA-liposome complexes. It was indicated that the transfection efficiency was the highest in the conditions of 1.0 μg DNA and 2.4 μL liposome within 24-pore plate, 85%-90% of the cell confluence and 4 h exposure time. The over doses of DNA and liposome could produce toxicity to the cells and impair the transfection effi- ciency, and the overtime exposure could also reduce the efficiency.

关 键 词：猪 耳成纤维细胞 绿色荧光蛋白基因 

分 类 号：S828[农业科学—畜牧学] S814.8[农业科学—畜牧兽医]