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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: To compare the different transfection characteristics of spiral ganglion cells (SGCs) in neonatal rats transfected with lentiviral vector and adenovirus vector. Methods: SGCs of neonatal rats were cultured in vitro and transfected into SGCs with appropriate titer of lentiviral vector and adenovirus vector. After 3 days (6 days after culture) and 7 days after transfection (10 days after culture), SGCs were transfected into SGCs. Fluorescence microscopy Green fluorescent protein reporter gene (GFP) expression and cell morphology of SGCs. The SGCs were counted, the transfection efficiency was calculated, and the length of axon of SGCs in each group was measured by ImageJ software. Results: The lentiviral vector and adenovirus vector could effectively transfect primary SGCs cultured in vitro. The adenovirus vector had high transfection efficiency and fast GFP expression, but the GFP expression began to decline after 1 week. After 7 days of transfection, GFP Began to express, and then transfected SGCs increased GFP enhanced. Compared with the control group, there was no significant difference in cell number and axon length between the SGC transfected with lentiviral vector or adenovirus vector. Conclusion: Both lentiviral vector and adenovirus vector can safely and effectively transfected primary cultured SGCs. Adenovirus vector transfection fast, efficient; lentiviral vector transfected SGCs GFP maintain a long time.