恒河猴大脑皮质星形胶质细胞的原代培养及鉴定  

Primary Culture and Identification of Astrocytes from Cereral Cortex of Rhesus Macaque

作　　者：王建斌 冯敏 廖芸 王丽春[1] 张莹 范胜涛 李琦涵[1] Wang Jianbin Feng Min Liao Yun Wang Lichunl Zhang Ying Fan Shengtao Li Qihan(Institute of Medical Biology, Chinese Academy of Medical Scienees and Peking Union Medical College, Yunnan Key Laboratory of Vaecine ReSearch and Development on Severe Infectious Diseases, Kunming, 650118, China Institute of Pathogen Biology Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100730, China)

机构地区：[1]中国医学科学院医学生物学研究所&北京协和医学院,云南省重大传染病疫苗研发重点实验室,昆明650118 [2]中国医学科学院病原生物学研究所&北京协和医学院,北京100730

出　　处：《中国细胞生物学学报》2017年第1期58-64,共7页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(批准号:31370192);中央高校基本科研业务费专项资金(批准号:2016ZX310047、2016ZX310179-2);中国医学科学院医学与健康科技创新工程重大协同创新项目(批准号:2016-I2M-1-019);云南省应用基础研究项目(批准号:2013FA024、2014FB191、2014HB066)资助的课题~~

摘　　要：为建立有效纯度高的恒河猴星形胶质细胞体外培养体系,无菌条件下取6月龄恒河猴婴猴的大脑皮质,除去白质,充分剪碎后机械吹打制成细胞悬液接种培养,待原代培养的细胞长满培养皿后,通过恒温摇床振荡和传代差速贴壁除去寡突胶质细胞、成纤维细胞等,得到纯化后的星形胶质细胞,并用GFAP-FITC(glial fibrillary acidic protein-fluorescein isothiocyanate)免疫荧光法对所获细胞进行鉴定。分离培养的细胞具备典型的星形胶质细胞形态,并表达星形胶质细胞特异性抗原GFAP(glial fibrillary acidic protein),纯化后获得了高纯度的恒河猴星形胶质细胞。采用该培养方法成功建立原代恒河猴星形胶质细胞培养体系,为体外研究嗜神经性病毒、神经系统疾病及其相关中枢神经病理机制等提供了可靠的细胞模型。The aim of this study is to establish an effective method to culture and identify the astrocytes from cerebral cortex of rhesus macaque. A male 6-month-old neonatal rhesus macaque was used, and the gray matter of cortex was isolated under aseptic conditions. The tissue was fully minced and pipetted to dispersed cell suspension to culture. The oligodendrocytes and microglias were removed through thermostatic oscillation, and differential attachment method was used to remove fibroblasts. The purified astrocytes were identified by immunocytochemical staining and immunofluorescence with anti-glial fibrillary acidic protein（GFAP）. The results showed that the purity of astrocytes of rhesus macaque exceeded 95%, and the morphology of these cells had the typical characteristics of astrocytes. Thus the method to culture highly purified astrocytes of rhesus macaque wasestablished successfully, which would be an invaluable tool for studying the roles of astrocytes in physiological and pathological states, and the pathogenesis and development of central nervous system diseases in vitro.

关 键 词：星形胶质细胞 恒河猴 原代培养 GFAP 

分 类 号：R338[医药卫生—人体生理学]