小鼠骨样细胞MLO-Y4转染方法的研究

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。

Study on transfection method to MLO-Y4 cells

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.