慢病毒转染海马神经元的模型建立  

Establishing model of lentivirus transfection of hippo-campal neurons

作　　者：于金鹏[1] 伍国锋[1] 王丽琨[1] 

机构地区：[1]贵州医科大学附属医院急诊神经内科,贵州贵阳550004

出　　处：《中风与神经疾病杂志》2016年第2期161-162,共2页Journal of Apoplexy and Nervous Diseases

摘　　要：目的建立一种简单易行的体外慢病毒转染海马神经元的实验方法。方法取怀孕的SD大鼠,分离体内胎鼠的海马后,经Accutase消化后接种,采用无血清培养海马神经元,选择合适的时间将慢病毒加入神经元内,于不同的时间在倒置荧光显微镜下观察细胞内GFP的显影效果。结果慢病毒转染海马神经元后,在第3天时观察,细胞内GFP在倒置荧光显微镜下显影达到80%以上。结论此方法简单易行,慢病毒转染神经元的成功率高,为慢病毒转染海马神经元后的后续实验奠定了实验基础。Objective To establish a simple and practical method of slow virus in vitro transfection method of hippocampal neurons. Methods The hippocampus were isolated from neonatal rat（ 1 d）,and digested with accutase. Hippocampal neurons were planted and cultuted with serum-free neurobassal. The Lentivirus were added into the neurons at the appropriate time,then observe the enhancement effect of GFP in the cell at different times In the inverted fluorescence microscope. Results Hippocampal neuron cultures are infected after 3 days in vitro by simply adding the amount of virus,and estimated to infect up to 80% of all neurons based on checking for fluorescent protein expression In the inverted fluorescence microscope. Conclusions The persent protocol is a simple and efficient method for lentiviral transfection of hippocampal neurons.

关 键 词：海马 神经元 慢病毒 转染 

分 类 号：R741[医药卫生—神经病学与精神病学]