胚胎大鼠神经干细胞体外分化的激光共聚焦显微镜观察  

Identification of Differentiation of Neural Stem Cells from Fetal Rats by Laser Scanning Confocal Microscopy

作　　者：任大林[1] 黄国伟[1] 张绪梅[1] 刘欢[1] 

机构地区：[1]天津医科大学公共卫生学院,300070

出　　处：《天津医药》2008年第10期777-779,I0008,共4页Tianjin Medical Journal

基　　金：国家自然科学基金资助项目(项目编号:30571563;30771797)

摘　　要：目的:采用激光扫描共聚焦显微镜观察体外培养胎鼠大脑皮质神经干细胞(neural stem cells,NSCs)分化情况。方法:利用无血清培养方法,分离培养胚胎大鼠大脑NSCs,进行体外扩增培养、传代;采用溴脱氧尿嘧啶核苷(bromodeoxyuridine,BrdU)掺入、双重免疫荧光细胞化学标记方法和激光扫描共聚焦显微镜,用神经细胞的特异性抗体(神经元β-微管蛋白、胶质纤维酸性蛋白),鉴定NSCs向神经元与星形胶质细胞分化的情况。结果:从胎鼠大脑皮质及皮质下分离的组织,经原代和传代培养均可形成细胞克隆,并表达神经上皮干细胞蛋白(Nestin)抗原。在血清诱导下,分化后的细胞表达神经元、星形胶质细胞2种神经细胞的特异性抗原,NSCs分化为星形胶质细胞神经元的比例分别为(43.70±8.55)%和(23.00±3.69)%。结论:从胎鼠大脑皮质分离出的细胞可获得呈集落样生长的神经干细胞团,并能表达NSCs的特异性抗原;激光扫描共聚焦显微镜可清晰地观察到培养细胞具有分化为神经元和星形胶质细胞的潜能。Objective： To observe the differentiation of neural stem cells （NSCs） originated from the brain cortex of embryonic Sprague-Dawley （SD） rat by laser scanning confocal microscopy. Methods： NSCs were isolated and cultured by microdisseetion, mechanical blowing and serum-free suspension culture, and identified by immunofluorescent staining using antibody against nestin. The differentiation of NSCs were identified by double staining immunofluorescence technique and laser scanning eonfocal microscope. Results： Neurosperes formed by NSCs were Nestin positive, and furthermore differentiated into neuron specific β-tubulin m positive cells and glial fibrillary acidic protein positive cells respectively. Ratio of astrocytes in differentiated cells was （43.70±8.55）%, and neurons was（23.00±3.69）% （P 〈 0.05 ）. Conclusion： The isolated cells from fetal rats possessed the potential ability of mutipotent differentiation. The potential ability of differentiation of NSCs into neurons or gli＇al cells can be observed clearly by double immunofluorescence staining and laser scanning.

关 键 词：干细胞 大脑皮质 神经系统 胚胎 激光 显微镜检查 共焦 溴脱氧尿苷 大鼠 

分 类 号：R651.2[医药卫生—外科学] R329.2[医药卫生—临床医学]