超声辐照参数对体外基因转染效果的影响  

Impact of ultrasonic irradiation parameters on in vitro gene transfection

作　　者：丁尚伟[1] 张艳容[1] 孙振兴[1] 张培歌 黄晓宇[1] 谢明星[1] 吕清[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院超声影像科湖北省分子影像重点实验室,湖北武汉430022

出　　处：《中国介入影像与治疗学》2014年第5期306-310,共5页Chinese Journal of Interventional Imaging and Therapy

基　　金：国家自然科学基金青年基金项目(81000616)

摘　　要：目的探讨超声辐照参数对体外基因转染效果的影响及最佳转染条件。方法将293T细胞种植于24孔板内,24h后更换为转染液进行超声辐照,EGFP质粒浓度为1mg/ml,每孔加入15μg;超声强度分为1.5、2.0、2.5W/cm2,辐照时间分为30、45、60s,微泡浓度分为20%、30%、40%;每个参数相互组合并重复4次。转染后72h于荧光显微镜下观察细胞内绿色荧光蛋白的表达,以流式细胞仪检测荧光细胞比例,CCK-8检测细胞存活率,计算转染效率。结果超声声强、辐照时间、微泡浓度均对转染效率有显著影响。辐照条件为2.0 W/cm2、辐照时间45s、微泡浓30%时转染效率最高,可达(35.25±1.40)%。结论超声辐照参数共同影响转染效果,低条件下相互协同,高条件下相互抑制,选择合适的辐照参数是超声介导体外基因转染的重要因素。Objective To investigate the impact of ultrasonic irradiation parameters in gene transfection and optimal transfection conditions. Methods The 293T cells were cultured in 24-well plates for 24 h, and then eGFP gene were trans- fected by ultrasound. Different parameters such as acoustic power （1.5, 2.0, 2.5 W/cm2） , exposure time （30, 45, 60 s） and microbubbles concentration （20%, 30 %, 40 %） were used. Three days after transfection, fluorescence microscope was used to observe the green fluorescent proteins of 293T cells, and flow cytometer was used to measure the proportion of 293T cells with the expression of green fluorescent proteins, and the activity of 293T cells was assessed by CCK-8, then the efficiency of transfection was calculated. Results All parameters had significant impact on transfection. The best transfect efficiency was （35.25±1.40）% under the condition of 2.0 W/cm2 , 45 s, 30%. Conclusion Many parameters have impact on gene transfect efficiency. Optimal ultrasonic parameters can significantly enhance gene delivery.

关 键 词：超声辐照 基因转染 微泡 绿色荧光蛋白 

分 类 号：R445.1[医药卫生—影像医学与核医学]