【摘要】 为了建立质粒转染小鼠骨样细胞MLO-Y4的方法,分别采用阳离子脂质体法和电转染法将增强型绿色荧光蛋白(EGFP)质粒pEGFP-C1转染小鼠骨样细胞MLO-Y4,正常培养48h后检测并统计转染率和死亡率。结果显示,脂质体法转染,当质粒与脂质体比例为1∶4时,转染效率可达到(36.8±3.7)%,细胞死亡率为(18.4±1.9)%;电转染法转染,脉冲电压240 V,脉冲时间300μs,脉冲次数3次时,转染率最高,可达到(23.8±2.3)%,细胞死亡率为(14.1±1.1)%。而后MTT实验显示脂质体转染法相对于电转染法对MLO-Y4细胞的增殖有一定的抑制作用,但对后续实验研究影响不大。脂质体转染法转染小鼠骨样细胞MLO-Y4优于电转染法。更多还原

【Abstract】 To select and optimize the transfection condition,MLO-Y4,a murine osteocyte-like cell line,was transfected with plasmid pEGFP-C1 by lipofection and electrotransfection.Transfection efficiency and mortality rate were measured after 48h.And then measure cell proliferation was assayed by MTT.The results showed when the ratio of plasmid to cationic liposome is 1:5,the lipofection efficiency reached(36.8 3.7)%,the mortality rate is(18.4 1.9)%;when pulse voltage is 240 V and pulse time is 300 μs,the electrotransfection efficiency was(23.8±2.3)%,the mortality rate reached(14.1±1.1)%.Lipofection could inhibit cell proliferation,but it had little effect on subsequent experiments.Taken all factors into consideration,for MLO-Y4 cells,lipofection transfection is a better method than electrotransfection.更多还原