SD大鼠大脑皮层星形胶质细胞的体外原代培养Primary Culture of Cerebral Cortical Astrocytes from SD Rats in vitro
汤婷婷,官志忠,禹文峰
TANG Tingting,GUAN Zhizhong,YU Wenfeng
摘要(Abstract):
目的:建立原代培养的SD大鼠大脑皮层星形胶质细胞体系。方法:取新生24~48 h的SD大鼠,无菌操作取出大脑皮层,剪碎后用胰酶消化、机械吹打制成分散细胞悬液,网筛过滤,将细胞悬液接种培养7~9 d;置于摇床(37℃、250 r/min振摇6 h),弃掉含脱离细胞的细胞悬液,去除小胶质细胞及少突胶质细胞,细胞传代,进行GFAP细胞免疫荧光鉴定。结果:成功分离和培养原代星形胶质细胞,GFAP免疫荧光鉴定星形胶质细胞比例在95%以上。结论:采用该培养方法能够成功的建立原代SD大鼠大脑皮层星形胶质细胞体系,为研究神经系统疾病及其相关疾病提供了可靠的细胞模型。
Objective: To establish primary cultured SD rat cerebral cortical astrocyte system. Methods: Cerebral cortex of SD neonatal rat within 24h was taken with aseptic operation and was cut to pieces,digested by trypsin,and dispersed cell suspension was made by mechanical method and filtered. Cell suspension was cultured for 7 to 9 days and were shaken in orbital shaker( 37 ℃,250 r / min for 6 h),and water from cell suspension was discarded to remove microglia and oligodendrocytes. Passaged cells were identified by immunofluorescent with anti-GFAP antibody. Results: Astrocytes were isolated and cultured successfully,and their ratios were identified to be above 95%. Conclusions: Primary cultured SD rat cerebral cortical astrocyte system is successfully established,which provide a reliable research model for diseases of nervous system and related studies.
关键词(KeyWords):
大鼠,Sprague-Dawley;星形胶质细胞;大脑皮层;细胞培养,原代;模型,细胞
rats,Sprague-Dawley;astrocytes;cerebral cortex;cell culture,primitive;model,cell
基金项目(Foundation): 国家自然科学基金(81360199);; 教育部科学技术研究课题(213032A)
作者(Author):
汤婷婷,官志忠,禹文峰
TANG Tingting,GUAN Zhizhong,YU Wenfeng
DOI: 10.19367/j.cnki.1000-2707.2014.02.005
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