NURR1基因修饰C17.2神经干细胞移植治疗脑梗死  

NURR1 gene-modified C17.2 neural stem cell transplantation for treatment of cerebral infarction

作　　者：黎祥喷[1] 沈庆煜[1] 王艺东[1] 叶剑虹[1] 邢诒刚[1] 

机构地区：[1]中山大学附属第二医院神经内科,广东省广州市510120

出　　处：《中国组织工程研究与临床康复》2007年第24期4760-4763,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：广东省卫生厅科研基金资助(B2004047)~~

摘　　要：目的:比较C17.2神经干细胞和NURR1基因修饰的C17.2神经干细胞移植治疗对脑梗死模型鼠神经功能缺损的改善效果,观察细胞移植后与宿主脑组织的整合、存活、移行及分化情况。方法:实验于2005-06/12在中山大学附属第二医院林百欣实验中心完成。①实验材料:选取清洁级健康SD雄性大鼠78只,随机数字表法分为模型对照组、神经干细胞组、转基因神经干细胞组,26只/组。条件永生化神经干细胞系C17.2由哈佛大学儿童医院Even Synder教授惠赠。pAdeasy-NURR1重组复制缺陷性腺病毒由本实验室成功构建。Dil18荧光染料(Chemicon公司产品)。②实验方法:收集C17.2神经干细胞和pAdeasy-NURR1+C17.2神经干细胞,各分为两管,一管加入Dil18荧光示踪剂。各组大鼠均采用线栓法建立局灶性脑缺血/再灌注损伤模型,3d后进行细胞移植,以江湾Ⅱ型脑立体定向仪进行立体定位,选择缺血半暗区3个位点为移植区:前囟头侧1mm,旁开2mm,深度1.2mm;前囟尾侧3mm,旁开1.5mm,深度1.2mm;前囟尾侧6mm,旁开2mm,深度1.2mm。模型对照组每位点注射单纯培养液2μL;神经干细胞组取6只每位点移植Dil18标记的C17.2神经干细胞悬液2μL,剩余20只每位点移植C17.2神经干细胞悬液2μL;转基因神经干细胞组取6只每位点移植Dil18标记的pAdeasy-NURR1+C17.2神经干细胞悬液2μL,余20只每位点移植pAdeasy-NURR1+C17.2神经干细胞悬液2μL。③实验评估:各组大鼠分别在移植前1d及移植后1周、2周、1个月进行神经功能缺损评分。移植后1周、2周、1个月进行荧光示踪检查及神经元特异烯醇化酶免疫组化检测。移植后6个月行组织学检查。结果:移植后1周,模型对照组死亡1只,神经干细胞组死亡2组,转基因神经干细胞组死亡2只,均淘汰并予以补充。①神经功能缺损评分:移植前1d各组大鼠神经功能缺损评分基本相似(P>0.05)。移植后1周、2周、1个月神经干细胞组、转�AIM： To compare the amelioration of C17.2 neural stem cells （NSCs） and NURR1 plus C17.2 NSCs transplantation on neurologic impairment of the rat models of cerebral infarction. To observe the integration, survival, migration and differentiation of donor cells after transplantation. METHODS： The experiment was conducted at Linbaixin Experimental Center of Second Affiliated Hospital of Sun Yat-sen University from June toDecember 2005.①A total of 78 healthy male SD rats of clean grade were selected and randomly divided into model control group （group A）, C17.2 neural stem cell transplantation group （group B） and NURR1 plus C17.2 neural stem cell transplantation group （group C） with 26 in each group. Conditionally immortal NSCs line C17.2 was presented by professor Even Synder who worked at Children＇s Hospital of Harvard University. The recombinant replication-defective adenovirus pAdeasy-NURR1 has been successfully constructed in this laboratory. Dill8 fluorescent dye was bought from Chemicon Company. ② C17.2 cells and pAdeasy-NURR1 plus C17.2 cells were collected and divided into two tubes, and then the Dill8 fluorescent dye was added into one of them. Rat models of focal cerebral ischemia/reperfusion injury were set up by thread-embolism method. Three days later cell transplantation was performed. Stereotaxis was done with Jianghuang type Ⅱ cerebral stereotaxic apparatus. Three sites were chosen in ischemic periphery area as transplanted sites： anterior fontanel anterior as 1 mm, lateral as 2 mm, ventral as 1.2 ram; anterior fontanel caudal as 3 mm, lateral as 1.5 mm, ventral as 1.2 mm; anterior fontanel caudal as 6 mm, lateral as 2 mm, ventral as 1.2 mm. Group A received 2 μL simple culture fluid at each of the 3 sites. Six rats taken from group B were infused with 2 μL C17.2 NSCs suspension labeled Dil18 fluorescent dye at each sites, and the others （20 rats） were infused with 2 μL C17.2 NSCs suspension at each sites. Six rats taken from group C were infused with 2 μL pAdeasy

关 键 词：NURR1基因 移植 脑梗死 C17.2神经干细胞 

分 类 号：R394.2[医药卫生—医学遗传学]