本文選題：神經(jīng)干細(xì)胞 + 腦室下帶區(qū)��； 參考：《華東師范大學(xué)》2006年碩士論文

【摘要】：[目的] 研究神經(jīng)干細(xì)胞移植到成年大鼠紋狀體后，在宿主腦內(nèi)的存活、遷移、分化等行為，為腦內(nèi)細(xì)胞移植提供基礎(chǔ)實(shí)驗(yàn)證據(jù)；探討神經(jīng)干細(xì)胞移植對(duì)PD模型動(dòng)物的治療改善情況。 [方法] 實(shí)驗(yàn)從E14胎鼠腦室下帶區(qū)(subventricular zone，SVZ)分離培養(yǎng)神經(jīng)干細(xì)胞。采用神經(jīng)球培養(yǎng)方法，獲得大量的細(xì)胞克隆。分別用抗Nestin抗體、抗NF抗體、抗GFAP抗體和抗CNP抗體，對(duì)培養(yǎng)的細(xì)胞進(jìn)行干細(xì)胞特性和多分化潛能的免疫組織化學(xué)鑒定。 原代培養(yǎng)的神經(jīng)干細(xì)胞經(jīng)3次傳代培養(yǎng)之后，選擇生長(zhǎng)狀態(tài)良好的神經(jīng)干細(xì)胞用AAV_2-GFP感染標(biāo)記，把帶有GFP標(biāo)記的神經(jīng)干細(xì)胞移植到成年大鼠紋狀體中。于細(xì)胞移植后的45天、90天和120天時(shí)分別灌流固定實(shí)驗(yàn)大鼠，對(duì)固定后的大腦進(jìn)行矢狀冰凍切片，熒光顯微鏡下觀察并配合免疫組織化學(xué)方法，研究移植細(xì)胞在大鼠腦內(nèi)的存活、遷移、分化等行為。 向健康成年大鼠的前腦內(nèi)側(cè)束部位注射適量6-羥多巴胺(6-OHDA)，建立帕金森病的大鼠模型，，一個(gè)月后，用阿撲嗎啡誘導(dǎo)模型大鼠旋轉(zhuǎn)，符合行為學(xué)要求的視為PD模型建立。對(duì)成功建立帕金森病模型的大鼠，向其腦內(nèi)紋狀體部位進(jìn)行神經(jīng)干細(xì)胞移植，研究神經(jīng)干細(xì)胞移植對(duì)PD模型大鼠的治療改善情況。 [結(jié)果] (1) 從E14胎鼠的SVZ部位分離得到的細(xì)胞具有增殖和形成神經(jīng)球的能力，神經(jīng)球表達(dá)Nestin，神經(jīng)球分化后的細(xì)胞分別對(duì)NF、GFAP、CNP具有免疫陽(yáng)性反應(yīng)。 (2) AAV_2-GFP感染標(biāo)記的神經(jīng)干細(xì)胞在熒光顯微鏡下呈現(xiàn)綠色熒光，分化后的GFP標(biāo)記細(xì)胞從胞體到突起整個(gè)細(xì)胞都有綠色熒光標(biāo)記。 (3) 在神經(jīng)干細(xì)胞移植后的各時(shí)間點(diǎn)，宿主腦內(nèi)都有大量標(biāo)記細(xì)胞存在。 (4) 神經(jīng)干細(xì)胞移植后45天，細(xì)胞遷移表現(xiàn)出明顯的方向性。神經(jīng)干細(xì)胞移植后90天，多數(shù)細(xì)胞表現(xiàn)為朝向腹后側(cè)遷移，遷移出的最遠(yuǎn)距離為1.0mm。神經(jīng)干細(xì)胞移植后120天，遷移的細(xì)胞主要表現(xiàn)為兩條遷移路線：一條細(xì)胞沿弧形路線向背后
[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
【學(xué)位授予單位】：華東師范大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類號(hào)】：R329

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