超声介导质粒GFP转染小鼠H2K成肌细胞的实验研究

目的 探讨超声介导基因转染小鼠骨骼肌细胞的作用。方法 采用质粒GFP作为目的基因，超声(频率1MHz，脉冲波，工作周期20％)作用于H2K成肌细胞，分别使用两种空间时间峰值强度(0．5W／cm^2、1W／cm^2)，照射时间分别为10s、20s、30s、40s、50s、60s。流式细胞仪测定GFP阳性细胞率，台盼蓝染色测定细胞生存率。结果 较低能量超声(0．5W／cm^2)GFP阳性细胞率总体显著低于较高能量超声(1W／cm^2)，超声增强GFP转染H2K细胞的最佳条件为：空间时间峰值强度1W／cm^2，照射时间40～50s。超声作用并未明显增加细胞死亡率。结论 超声在增强骨骼肌细胞基因转染领域应用前景广阔。

Experiment of ultrasound-mediated plasmid GFP transfer into H2K cell in vitro

Objective To explore the effect of ultrasound to gene transfer in skeletal muscle cells in mice. Methods All comparisons were performed between two intensities of ultrasound((0.5) W/cm~2, (1 W/cm~2)) with pulse wave (1 MHz, 20% duty cycle) using plasmid green fluorescent protein(GFP) on H2K myoblast cells. Exposure duration was 10 s, 20 s, 30 s, 40 s, 50 s, and 60 s. The trend of transfection efficiency by flow cytometer and cell viability by Trypan blue assays under different conditions was measured. Results GFP transfection rate in H2K cell treated by (0.5) W/cm~2 was significantly lower than that by 1 W/cm~2 as a whole. The optimum condition of GFP transfection for H2K cell was 1 W/cm~2 and (40-)50 s. Ultrasound did not increase the rate of cell-killing. Conclusions Ultrasound holds promise in gene delivery in skeletal muscle cells.