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成年大鼠纹状体区神经干细胞的分离培养 |
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刘仕勇 张可成 杨辉 蔡文琴 何家全
摘 要：目的 从成年大鼠纹状体区分离并鉴定神经干细胞。 方法 利用无血清培养和单细胞克隆技术在成年大鼠纹状体区分离具有单细胞克隆能力的细胞群，选取单个克隆进行连续传代培养获得大量亚克隆，采用免疫荧光细胞化学技术检测神经上皮干细胞蛋白(Nestin)和分化后特异性成熟神经细胞抗原的表达。 结果 从纹状体区分离的细胞群具有连续克隆能力，表达神经上皮干细胞蛋白，分化后的细胞表达神经元、星形胶质细胞和少突胶质细胞的特异性抗原。 结论 分离的细胞具有自我更新能力和多分化潜能，表达神经上皮干细胞蛋白，有很强的增殖能力。是中枢神经系统的干细胞。
关键词：神经干细胞；克隆分析；细胞培养；分离；鉴定
分类号：Q813 文献标识码：A
文章编号：0529-1356(2000)01-17
ISOLATION OF NEURAL STEM CELLS FROM
STRIATUM OF ADULT RATS
LIU Shi-yong
(Department of Neurosurgery,Xinqiao Hospital)
ZHANG Ke-cheng
(Department of Neurosurgery,Xinqiao Hospital)
YANG Hui
(Department of Neurosurgery,Xinqiao Hospital)
CAI Wen-qing
(Department of Histology and Embryology,The Third Military Medical University,Chongqing 400037,China)
HE Jia-quan
(Department of Histology and Embryology,The Third Military Medical University,Chongqing 400037,China)
Abstract：Objective To isolate neural stem cells from striatum of adult rats.Methods A cell line which had ability to form clones was isolated from striatum of adult rats by using serum-free cell culture and single cell clone test.A clone was passaged continuously in order to acquire a large number of clones,and immunofluorescence was used to detect Nestin antigen of the clone and specific antigen of mature neural cell after the clone differentiated.Results The cell line isolated from striatum of adult rats expresses Nestin antigen and has the potential to form clones and differentiate into neurons,astrocytes,and oligodendrocytes.Conclusion Our cell line which expresses Nestin antigen is multipotent cell line and has ability to self-renew,and believed that it contains stem cells of the CNS.
Key words：Neural stem cell;Clonal assay;Cell culture;Isolation;Identification;▲
成年哺乳类动物神经系统通常被看作无再生能力的组织，意指神经细胞不能分裂增殖产生新生神经元以替代丢失、缺损的神经元。但近年来的研究对此提出了疑问。首先，人们发现成年个体海马和室管膜下层细胞具有分裂产生新生神经元的能力，尤其是位于室管膜下层的祖细胞增殖后可以沿着一条比较固定[1] [2] 下一页
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