慢病毒介导CD-TK融合基因转染对神经干细胞体外增殖的影响  

Effect of CD-TK fusion gene transfection on the proliferation of neural stem cells in vitro

作　　者：邢春阳[1] 牛健[1] 蒋传路[1] 杨海城[1] 

机构地区：[1]哈尔滨医科大学附属第二医院神经外科,150086

出　　处：《中国微侵袭神经外科杂志》2013年第4期175-177,共3页Chinese Journal of Minimally Invasive Neurosurgery

基　　金：黑龙江省自然科学基金项目(编号:D2007-31);中国博士后科学自助基金(编号:20080430138);黑龙江省教育厅科学技术研究项目(编号:11531185)

摘　　要：目的探讨慢病毒介导胞嘧啶脱氨酶(cytosine deaminase,CD)及胸腺嘧啶激酶(thymidine kinase,TK)融合基因转染对神经干细胞体外增殖的影响。方法原代培养新生Wistar大鼠室下区神经干细胞并鉴定,经传代纯化后,用CD-TK基因包装的慢病毒处理3～5 d。MTT实验检测神经干细胞增殖活性,免疫荧光技术检测转染后神经干细胞GFAP、NSE表达。结果 MTT结果显示CD-TK融合基因转染对神经干细胞增殖无影响(P>0.05)。免疫荧光结果显示:神经干细胞Nestin标记阳性;CD-TK融合基因转染后,神经干细胞NSE及GFAP均表达阳性。结论 CD-TK融合基因转染对神经干细胞增殖无影响。Objective To explore the effects of lentivirus-mediated cytosine deaminase（CD）-thymidine kinase（TK） fusion gene transfection on the proliferation of neural stem cells（NSCs） in vitro.Methods Primary NSCs obtained from the subventricular zone（SVZ） of newborn Wistar rats were cultured and identified.After passage and purification in vitro,NSCs were treated with lentiviruses and packaged with CD-TK gene for 3 to 5 d,and then MTT assay was used to detect the proliferation of NSCs.Immunofluorescence technique was used to detect the expressions of glial fibrillary acidic protein（GFAP） and neuronspecific enolase（NSE） in NSCs.Results CD-TK gene transfection showed no influence on the proliferation of the NSCs（P ＆gt; 0.05）.The NSCs was positive for nestin and the naturally differentiated cells were positive for GFAP and NSE by immunofluorescence after incomplete culture.Conclusion CD-TK gene transfection does not affect proliferation of NSCs.

关 键 词：神经干细胞 CD-TK融合基因 慢病毒介导 体外培养 细胞增殖 大鼠 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]