成年大鼠纹状体神经干细胞的体外培养及分化  

In vitro culture and induced differentiation of adult rat neural stem cells from the corpus striatum

作　　者：王彦惠[1] 刘亚杰[1] 卢洪流[2] 刘振华[1] 姜晓丹[2] 徐如祥[2] 周振军[2] 邹雨汐[2] 陈一招[2] 

机构地区：[1]第一军医大学珠江医院神经内科,广东广州510282 [2]第一军医大学珠江医院神经外科,广东广州510282

出　　处：《第一军医大学学报》2004年第2期192-194,197,共4页Journal of First Military Medical University

基　　金：军队科技项目(01Z054);广东省重大科技项目[粤科基办(2000)25] ~~

摘　　要：目的探讨成年大鼠纹状体神经干细胞多向分化的可能性。方法分离成年大鼠纹状体神经干细胞,单克隆连续传代培养并诱导分化,将分化产物进行免疫细胞化学染色及RT-PCR检测分析。结果无血清单克隆培养时期可见大量的细胞团出现,以血清诱导分化后出现多种细胞类型,免疫细胞染色发现分化细胞中有Nestin阳性、NSE阳性、GFAP阳性细胞,RT-PCR法分析mRNA水平有脑因子-1、γ-氨基丁酸α-受体γ-亚单位、酪氨酸羟化酶及色氨酸羟化酶等基因的表达。结论成年大鼠纹状体内分离出的干细胞样细胞具有自我增殖和多向分化潜能。Objective To investigate the in vitro multipotential differentiation of neural stem cells from adult rat corpus striatum. Methods The neural stem cells isolated from adult rat corpus striatum were cultured in serum-free medium to obtain cell suspension before monoclonal subculturing and differential induction. Immunocytochemical staining and reverse transcriptional PCR (RT-PCR) were performed to identify the properties of the differentiated cells. Results Numerous cell clusters were formed in the phase of monoclonal culture, and different types of cells were observed 3 d after induction with fetal bovine serum. The differentiated cells contained cells positive for nestin, neuron-specific enolase (NSE) positive cells, and glial fibrillary acidic protein (GFAP) positive cells. RT-PCR identified expressions of the transcripts for neural cell-associated genes including brain factor-1, γ-aminobutyric acid α-receptor γ-subunit, tyrosine hydroxylase and tryptophan hydroxylase. Conclusion The cells separated from adult rat corpus striatum possess the ability of self-proliferation and multipotential differentiation, and are identified as the stem cells of the central nervous system.

关 键 词：成年大鼠 纹状体 神经干细胞 体外培养 细胞分化 逆转录聚合酶链反应 细胞培养 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] R-332[医药卫生—基础医学]