慢病毒载体和腺病毒载体转染原代培养的螺旋神经节细胞的比较  

Comparison of lentivirus and adenovirus vector mediated gene transfer into cultured spiral ganglion cells

作　　者：王国鹏[1] 罗凌惠[2] 谢静[1] 陈沛[2] 付勇[3] 钟刚[2] 龚树生[1] 

机构地区：[1]首都医科大学附属北京同仁医院耳鼻咽喉头颈外科,北京100730 [2]华中科技大学同济医学院附属协和医院耳鼻咽喉科 [3]浙江大学医学院附属第一医院耳鼻咽喉科

出　　处：《临床耳鼻咽喉头颈外科杂志》2011年第4期172-175,共4页Journal of Clinical Otorhinolaryngology Head And Neck Surgery

基　　金：国家自然科学基金(No:30872858);北京市自然科学基金(No:7082024);北京市教育委员会科技发展计划项目(No:KM200810025004)

摘　　要：目的:比较慢病毒载体和腺病毒载体转染新生大鼠耳蜗螺旋神经节细胞(SGCs)的不同转染特性。方法:体外培养新生大鼠SGCs,适当滴度的慢病毒载体和腺病毒载体转染SGCs,于病毒转染后3d(培养后6d)和转染后7d(培养后10d),荧光显微镜下观察绿色荧光蛋白报告基因(GFP)表达情况和SGCs的细胞形态。行SGCs计数,计算转染效率,并通过ImageJ软件测量各组SGCs轴突长度。结果:慢病毒载体和腺病毒载体均可有效转染体外培养的原代SGCs;腺病毒载体转染效率高,GFP表达快,而1周后GFP表达开始下降;慢病毒载体转染7d后GFP开始表达,而后被转染的SGCs增多,GFP增强。慢病毒载体或腺病毒载体转染后的SGC与对照组相比,细胞数目和轴突长度的差异均无统计学意义。结论:慢病毒载体和腺病毒载体均可安全、有效地转染原代培养的SGCs。腺病毒载体转染快,效率高;慢病毒载体转染SGCs后GFP维持时间长。Objective：We aimed to compare the characteristics between lentivirus and adenovirus vector mediated gene transfer into cultured spiral ganglion cells（SGCs）.Method：SGCs from newborn rats were cultured and exposed to lentivirus-GFP and adenovirus-GFP vectors.GFP expression and the cell morphology were evaluated under epi-fluorescence microscope at 3 days and 7 days after exposure.Survival number of SGCs was counted,and the average percentage of SGCs with GFP expression was calculated,and axon length was measured by ImageJ software.Result：Cultured SGCs were transfected by either adenovirus or lentivirus vector successfully.The adenovirus vector presented an instant and efficient transfection.However,the expression of GFP went down after 7 days.In lentivirus-GFP group,GFP expression was detected at 7 days after exposure,and the number of cells with GFP expression increased gradually in the following days.Statistical analysis revealed that there were no differences in survival number of SGCs and average axon length among lentivirus-GFP group,adenovirus-GFP group and control group.Conclusion：Cultured SGCs can be transfected by either lentivirus vector or adenovirus vector safely and efficiently.SGCs are more susceptible to adenovirus vector,but GFP persists for a longer period after the lentivirus-mediated gene transfer.

关 键 词：听觉丧失 感音神经性 慢病毒载体 腺病毒载体 基因转染 原代细胞培养 螺旋神经节细胞 

分 类 号：R764.43[医药卫生—耳鼻咽喉科]