【作者】 赵慧英； 温叶飞； 曹洪国； 华松； 尹宝英； 张涌；

【Author】 ZHAO Hui-ying, WEN Ye-fei, CAO Hong-guo, HUA Song, YIN Bao-ying, ZHAN Yong(Institute of Biotechnology, Northwest Sci-Tech University of Agriculture and Forestry, Wangling, Shaanxi, 712100,China)

【机构】 西北农林科技大学生物工程研究所；

【摘要】 为了进一步完善神经干细胞(Neural Stem Cells,NSCs)体外分离培养和诱导分化体系,本实验选取胚龄E14～15的小鼠胚胎,对胎鼠中枢神经系统(Central Nervous System,CNS)大脑皮质NSCs采用机械分离法制成单细胞悬液,在NSCs无血清生长培养液中进行原代和传代培养,在分化培养液和纹状体提取液中分别进行自然分化和诱导分化培养;并用免疫组织化学法对神经球及分化细胞进行鉴定。结果:在培养的第2～3d即有悬浮生长的神经球形成,并逐渐变大,每5d～7d传代1次可传至12代;细胞克隆率可达到95%;原代及传代生长的神经球均呈NSCs标志性蛋白-Nestin阳性;神经球或单细胞贴壁后可分化为神经元样细胞和星形胶质样细胞形态:由纹状体提取液组成的多巴胺能神经元诱导液可使14%的NSCs表达TH。证明分离得到具有干细胞特征和多分化潜能的NSCs。更多还原

【Abstract】 In order to establish the cell culture and differentiation system, the paper using serum free culture method isolate directly and culture central nervous system stem cells from embryonic day 14-15 of mouse cerebra. The differentiating medium are used to induced the neurospheres and single NSCs differentiation. Striatal extracts are used to induced NSCs to differentiate into dopaminergic neurons. The immunohistochemistry method are used to detect Nestin, NF and GFAP antigen of cells. The results are as follow: The cells isolated from fetal mouse cerebral expresses nestin antigen. They have potential to serially passage 12 th and differentiate into neurons and glial. Single cells from neurosphere cultures initiated from cells were again replaced as single cells and were able to reestablish neurosphere cultures, which clone ratio reach to 95%. Striatal extracts could induce 14% of them to differentiate into dopaminergic neurons. These indicate that the cells, which express nestin antigen, isolate from cerebral are multipotent and have the potential of self-renewal, therefore they are neural stem cells. The stem cells provide materials for engraftment, proliferation, migration, an d neural differentiation.更多还原