神经干细胞纹状体内移植后的行为和功能研究

发布时间：2018-06-08 12:21

本文选题：神经干细胞 + 脑室下带区　；参考：《华东师范大学》2006年硕士论文

【摘要】：[目的] 研究神经干细胞移植到成年大鼠纹状体后，在宿主脑内的存活、迁移、分化等行为，为脑内细胞移植提供基础实验证据；探讨神经干细胞移植对PD模型动物的治疗改善情况。 [方法] 实验从E14胎鼠脑室下带区(subventricular zone，SVZ)分离培养神经干细胞。采用神经球培养方法，获得大量的细胞克隆。分别用抗Nestin抗体、抗NF抗体、抗GFAP抗体和抗CNP抗体，对培养的细胞进行干细胞特性和多分化潜能的免疫组织化学鉴定。原代培养的神经干细胞经3次传代培养之后，选择生长状态良好的神经干细胞用AAV_2-GFP感染标记，把带有GFP标记的神经干细胞移植到成年大鼠纹状体中。于细胞移植后的45天、90天和120天时分别灌流固定实验大鼠，对固定后的大脑进行矢状冰冻切片，荧光显微镜下观察并配合免疫组织化学方法，研究移植细胞在大鼠脑内的存活、迁移、分化等行为。向健康成年大鼠的前脑内侧束部位注射适量6-羟多巴胺(6-OHDA)，建立帕金森病的大鼠模型，，一个月后，用阿扑吗啡诱导模型大鼠旋转，符合行为学要求的视为PD模型建立。对成功建立帕金森病模型的大鼠，向其脑内纹状体部位进行神经干细胞移植，研究神经干细胞移植对PD模型大鼠的治疗改善情况。 [结果] (1) 从E14胎鼠的SVZ部位分离得到的细胞具有增殖和形成神经球的能力，神经球表达Nestin，神经球分化后的细胞分别对NF、GFAP、CNP具有免疫阳性反应。 (2) AAV_2-GFP感染标记的神经干细胞在荧光显微镜下呈现绿色荧光，分化后的GFP标记细胞从胞体到突起整个细胞都有绿色荧光标记。 (3) 在神经干细胞移植后的各时间点，宿主脑内都有大量标记细胞存在。 (4) 神经干细胞移植后45天，细胞迁移表现出明显的方向性。神经干细胞移植后90天，多数细胞表现为朝向腹后侧迁移，迁移出的最远距离为1.0mm。神经干细胞移植后120天，迁移的细胞主要表现为两条迁移路线：一条细胞沿弧形路线向背后
[Abstract]:[objective] to study the survival, migration and differentiation of neural stem cells (NSCs) in the host brain of adult rat striatum after transplantation, so as to provide basic experimental evidence for brain cell transplantation. To investigate the therapeutic effect of neural stem cell transplantation on PD model animal. [methods] Neural stem cells were isolated and cultured from subventricular zone of E14 fetal rat. A large number of cell clones were obtained by using the method of neurosphere culture. Anti nestin antibody, anti NF antibody, anti GFAP antibody and anti CNP antibody were used to identify the stem cell characteristics and multi differentiation potential of cultured cells respectively. Neural stem cells (NSCs) in good growth state were selected to be labeled with AAVS 2-GFP infection and then transplanted into the striatum of adult rats with GFP labeled NSCs. After 45 days, 90 days and 120 days after cell transplantation, the experimental rats were perfused respectively. The sagittal frozen sections of the fixed brain were observed under fluorescence microscope and the survival of transplanted cells in the rat brain was studied with immunohistochemical method. The rat model of Parkinson's disease was established by injecting appropriate amount of 6-hydroxydopamine 6-OHDAA into the medial forebrain tract of healthy adult rats. After one month, the rat model was rotated with apomorphine. According to behavioral requirements, PD model is established. Neural stem cells (NSCs) were transplanted into the striatum of the rat model of Parkinson's disease. To study the effect of neural stem cell transplantation on the treatment of PD model rats. [results] the cells isolated from SVZ of fetal E14 mice have the ability to proliferate and form neurospheres. Nestin was expressed in the neurospheres, and the cells differentiated from the neurospheres showed positive immunoreactivity to NFFP-GFAPP CNP respectively. (2) the neural stem cells labeled with AAV2-GFP infection showed green fluorescence under fluorescence microscope. The GFP labeled cells had green fluorescent labeling from the cell body to the process. 3) at every time point after neural stem cell transplantation, there were a large number of labeled cells in the host brain. 4) 45 days after neural stem cell transplantation, there were many labeled cells in the host brain. Cell migration showed obvious directionality. 90 days after NSCs transplantation, most of the cells migrated toward the ventral posterior side, and the longest migration distance was 1.0 mm. 120 days after transplantation of neural stem cells, the migrating cells showed two main migration routes: one cell along the arc route to the back
【学位授予单位】：华东师范大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R329

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