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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/cm2、1W/cm2),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/cm2)GFP阳性细胞率总体显著低于较高能量超声(1W/cm2),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/cm2,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。
Objective To investigate the role of ultrasound-mediated gene transfection in mouse skeletal muscle cells. Methods Plasmid GFP was used as the target gene. Two kinds of time-dependent peak intensities (0.5W / cm2, 1W / cm2) were applied to the H2K myoblasts under ultrasound (frequency 1MHz, pulse wave and duty cycle 20% For 10s, 20s, 30s, 40s, 50s, 60s. The rate of GFP positive cells was determined by flow cytometry and the cell survival rate was determined by trypan blue staining. Results The rate of GFP-positive cells in low-energy ultrasound (0.5W / cm2) was significantly lower than that in higher-energy ultrasound (1W / cm2). The optimal conditions of enhanced GFP- Irradiation time 40 ~ 50s. Ultrasound did not significantly increase cell death rate. Conclusion Ultrasound in the field of enhancing gene transfer of skeletal muscle cells has broad prospects for application.