GFP转基因鼠神经上皮干细胞纹状体内移植的实验研究  

Survival and differentiation of GFP transgenic mice-derived neuroepithelial cells after being transplanted into rat brains

作　　者：王家增[1] 暴丽华[1] 孙晋浩[1] 李振华[1] 张静[1] 

机构地区：[1]山东大学医学院人体解剖学教研室,山东济南250012

出　　处：《山东大学学报（医学版）》2007年第7期649-654,共6页Journal of Shandong University:Health Sciences

基　　金：山东省优秀中青年科学家科研奖励基金(2004BS02012);山东省卫生厅科研资助基金(05210212)

摘　　要：目的:观察GFP转基因鼠胚胎神经上皮干细胞的体外培养及脑内移植后的存活与分化状况。方法:将GFP转基因鼠的胚胎神经上皮干细胞置于无血清培养基内纯化扩增,用巢蛋白免疫组织化学染色鉴定神经干细胞,将扩增后的含GFP基因的神经干细胞在立体定位仪下,定向移植到大鼠的纹状体内。术后不同时期取材、切片,在荧光显微镜下观察移植细胞的存活状况,行微管相关蛋白(MAP-2)、胶质纤维酸性蛋白(GFAP)以及酪氨酸羟化酶(TH)免疫组织化学染色鉴定移植细胞的分化状况。结果:GFP转基因鼠神经管来源的神经上皮干细胞在无血清培养基内能增殖形成神经球,呈巢蛋白阳性。纹状体内移植2周、4周均可见明显的呈绿色荧光的移植细胞,显示细胞存活良好。免疫组织化学染色显示移植细胞在不同间隔时间可分化为MAP-2、GFAP及TH阳性细胞。结论:GFP转基因鼠来源的具有GFP标记的胚胎神经上皮干细胞可在体外培养扩增,移植到大鼠纹状体内存活良好并能分化成神经元、神经胶质细胞,且能定向分化为多巴胺能神经元。Objective： To observe the survival and differentiation of ceils derived from the embryonic neural tube of green fluorescent protein （GFP） transgenic mice after being transplanted into the striatum of rats. Methods： Neuroepithelial stem ceils derived from the embryonic neural tube of GFP transgenic mice were cultured and depurated in free serum medium containing B27, bFGF and EGF, and then they were directionally transplanted into the striatum of rats. Animals were sacrificed at different time points after being transplanted and frozen sectioned, and staining by MAP- 2, GFAP, and TH was determined by immunohistochemical techniques. Results： Neuroepithelial stem ceils derived from the embryonic neural tube could expand into neurospheres in the free serum medium and express nestin. The grafted regions were clearly visible, and the transplanted ceils survived well and some of them differentiated into TH-positive cells. Condusion： Embryonic neuroepithelial stem ceils from GFP transgenic mice could be generated in vitro, survive and differentiate into neurons, especially dopaminergic neurons and neuroglia ceils after being transplanted into rat brains.

关 键 词：小鼠 转基因 神经上皮 干细胞 增殖 移植 

分 类 号：R322.81[医药卫生—人体解剖和组织胚胎学]