新生大鼠海马源性神经干细胞的分离培养及其向胆碱能神经元定向诱导分化研究(英文)  

ISOLATION OF NEURAL STEM CELLS FROM NEONATAL RAT HIPPOCAMPUS AND THEIR IN VITR O DIFFERENTIATION INTO CHOLINERGIC NEURONS

作　　者：罗湘颖[1] 杨志敏[2] 宋晓斌[2] 刘苏[1] 赵匡彦[2] 冯忠堂[1] 王廷华[1] 

机构地区：[1]昆明医学院神经科学研究所,昆明650031 [2]昆明医学院第一附属医院神经外科,昆明650032

出　　处：《神经解剖学杂志》2005年第2期190-194,共5页Chinese Journal of Neuroanatomy

基　　金：国家自然科学基金(No. 30260112);云南省自然科学基金(No. 2000C11R)资助项目

摘　　要：本研究目的是从新生SD大鼠海马分离、培养神经干细胞并诱导其向胆碱能神经元方向分化。利用含b FGF(20ng/ml)和B27的无血清DMEM/F12培养基培养新生SD大鼠海马分离的具有自我更新和多向分化能力的细胞群,用免疫细胞化学技术检测巢蛋白(nestin),并于分化后分别检查特异性成熟神经细胞、星形胶质细胞、少突胶质细胞的标记抗原β微管蛋白(Tuj1 )、胶质纤维酸性蛋白(GFAP)和半乳糖脑苷脂(Galc)的表达;用鸡胚骨骼肌提取液,诱导神经干细胞向胆碱能神经元方向分化。结果显示:从海马分离的细胞群具有自我更新能力,表达nestin,分化成熟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;与对照组3. 9%相比,鸡胚骨骼肌提取液可以诱导这些细胞中的9. 6%分化成为胆碱能神经元。提示分离的细胞具有自我更新能力和多向分化潜能,是中枢神经系统的干细胞;在加有鸡胚骨骼肌提取液的培养基诱导下,能向胆碱能神经元方向分化。The present study aims to isolate neural stem cells from neonatal rat hippocampus and induce them to differentiate into cholinergic neurons. A multipotent cell line derived from the hippocampi of neonatal rats which had the ability to form clones was incubated in serum-free DMEM/F12 medium added with 20ng/ml basic fibroblast growth factor (bFGF) and B27. After differentiation of the neural stem cells, immunocytochemistry was used to detect nestin, the antigen of the cell clone, and β-tubulin (Tuj1), glial fibrillary acidic protein (GFAP) and galactocerebroside (Galc), the markers specific for neurons, astrocytes and oligodendrocytes, respectively. Embryonic chick skeletal muscle extract was used to induce the differentiation of the neural stem cells into cholinergic neurons. The results showed that the cell line isolated from the hippocampi of neonatal rats expressed nestin and had the potential to form clones and differentiate into neurons, astrocytes and oligodendrocytes. Embryonic chick skeletal muscle extract can induce 9.6% of the isolated cell line to differentiate into cholinergic neurons compared with 3.9% in controls. These findings suggested that the cell line, which expressed nestin antigen, was a multipotent cell line capable of self-renewing, and was believed to contain stem cells of the CNS. These neural stem cells can be induced to differentiate into cholinergic neurons by using embryonic chick skeletal muscle extract.

关 键 词：大鼠 海马源性神经干细胞 细胞培养 胆碱能神经元 诱导 分化 免疫细胞化学 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]