中脑神经前体细胞的体外分离、培养和特性  

ISOLATION,CULTURE AND CHARACTERIZATION OF MESENCEPHALIC PROGENITOR CELLS IN VITRO

作　　者：孙秀[1] 陈生弟[1] 刘振国[1] 刘卫国[1] 梁粱[1] 徐洁懿[1] 

机构地区：[1]上海第二医科大学瑞金医院神经内科,上海200025

出　　处：《细胞生物学杂志》2002年第5期295-299,共5页Chinese Journal of Cell Biology

基　　金：国家重点基础研究规划"脑功能和脑重大疾病的基础研究";国家自然科学基金(G19999054008);国家自然科学基金(39970263);上海市卫生系统百名跨世纪优秀学科带头人培养计划(97BR001);上海二医大-上海生命科学中心科研项目

摘　　要：为了解中脑神经前体细胞的体外培养特性和建立中脑神经前体细胞的体外分化调控机制提供细胞模型。本实验采用含有丝分裂源表皮生长因子(EGF)的无血清培养基培养来源于大鼠胚胎E14.5天的中脑神经前体细胞,应用免疫细胞化学方法了解其前体细胞特性。结果发现中脑神经前体细胞呈神经前体细胞特征性标记Nestin免疫染色阳性,无分化细胞标记;细胞克隆实验证实中脑神经前体细 胞有自我更新能力;在EGF刺激下增殖迅速;当撤去EGF后置于含胎牛血清的培养基和被覆多聚赖氨酸(PLL)的培养皿内,中脑神经前体细胞可分化成神经元和星形胶质细胞。本试验证明我们培养的中脑神经前体细胞具有增殖、自我更新能力和多向分化潜能特性。To understand the characterization of neural progenitor cells in vitro and provide a cell model for studying differentiation of mesen-cephalic progenitor cells. The rat embryonic E14.5 mesencephalic progenitor cells were isolated and cultured in serum-free medium containing the mitogen epidermal growth factor(EGF). The characterization of progenitor cells was investigated by immunocytochemistry. The cultured mesencephalic cells were positively immunoreactive for progenitor cell marker Nestin, immunonegative for differentiated cell markers, and remained mitotically active. Clonal and population analyses showed their extensive self-renew capacity. The cells are EGF reactive,i, e. proliferate quickly in the medium containing EGF. When growing on poly-1-lycine coated plastic in serum contained medium, the mesencephalic progenitor cells spontaneously differentiate into both neurons and astroglia cells. Our research showed the cultured mesencephalic cells preserved critical characteristics of progenitor cell,including proliferation,self-renew and multipotency.

关 键 词：中脑 神经前体细胞 体外分离 细胞培养 特性 

分 类 号：Q813[生物学—生物工程]