麦胚凝集素慢病毒载体的构建及其对脂肪干细胞的感染  

Construction of a lentivirus vector expressing wheat germ agglutinin and its infection in human adipose-derived stem cells

作　　者：王娟[1] 张连国[2,3] 李宁[4] 雷鸣[1] 马云[2,5] 彭媛红 卞伟华[1] WANG Juan ZHANG Lianguo LEI Ming MA Yun BIAN Weihua(School of Pharmaceutical Sciences, Binzhou Medical College, Yantai 264003, China Department of Thoracic Surgery Department of Central Laboratory Binzhou Medical College Hospital, Binzhou 256600, China Department of obstetrics, Binzhou Medical College Hospital, Yantai 264003, China Shanghai Jiaotong University Hospital, Shanghai 200025, China)

机构地区：[1]滨州医学院药学院,山东烟台264003 [2]滨州医学院附属医院,山东滨州256600 [3]滨州医学院附属医院胸外科,山东滨州256600 [4]滨州医学院附属医院妇产科,山东烟台264003 [5]滨州医学院附属医院中心实验室,山东滨州256600 [6]上海交通大学医学院,上海200025

出　　处：《南方医科大学学报》2016年第9期1242-1246,共5页Journal of Southern Medical University

基　　金：国家自然科学基金(31401258);山东省自然科学基金(ZR2012BM006,ZR2013CL004)

摘　　要：目的探讨麦胚凝集素(WGA)慢病毒载体的构建与表达的方法,以及WGA在神经系统中跨膜示踪的能力。方法采用基因工程方法,将WGA的基因插入到慢病毒表达载体Plvx-IRES-Zs Green1中。再采用慢病毒包装,将构建好的质粒和其他3种包装质粒(RPEV、PRRE、VSVG)在293T细胞中进行共转染,获取病毒液后感染人源的脂肪干细胞(h ADSCs),将感染后的h ADSCs注入小鼠的脑卒中模型,修复神经损伤。结果显微镜下检测到绿色荧光报告蛋白表示感染成功。对感染后的h ADSCs进行免疫组化显色,结果显示WGA已经成功的在h ADSCs中实现了表达,在MCAO小鼠模型脑损伤处追踪到感染了WGA的h ADSCs。结论慢病毒包装WGA蛋白的方法可靠高效,可以作为一种高效的示踪技术广泛应用于各类细胞。Objective To construct a lentivirus vector carrying wheat germ agglutinin （WGA） and evaluate its ability of tracing WGA in the brain of mice with ischemic brain injury. Methods WGA gene was inserted into the lentiviral vector Plvx IRES-ZsGreen1 using genetic engineering methods. 293T cells were transfected with the vector and 3 packaging plasmids （RPEV, PRRE, and VSVG） to obtain the recombinant lentivirus for infection of human adipose-derived stem cells （hADSCs）. The infected hADSCs were injected into the damaged brain area by in situ injection in a mouse model of middle cerebral artery occlusion （MCAO） and the expression of GFP was traced. Results Immunofluorescence identification detected WGA protein expression in the infected hADSCs, which survived in the infarct area of mice with MCAO. Conclusion Packaging WGA gene in lentivirus is a reliable approach to allow efficient neuroanatomical tracing of various cells.

关 键 词：麦胚凝集素 慢病毒载体 免疫荧光 人源脂肪干细胞 

分 类 号：R743[医药卫生—神经病学与精神病学] Q78[医药卫生—临床医学]