ICR小鼠神经干细胞的体外培养与鉴定  

In vitro culture and identification of neural stem cells in ICR mice

作　　者：孙孟军[1] 郭建美[2] 周更苏[3] 董泽飞[4] 郑春贵 曹翠丽[5] SUN Meng-jun;GUO Jian-mei;ZHOU Geng-su;DONG Ze-fei;ZHENG Chun-gui;CAO Cui-li(Department of Public Teaching,Xingtai Medical College,Hebei Province,Xingtai 054002,China;Department of Basic Medicine,Xingtai Medical College,Hebei Province,Xingtai 054002,China;Department of Nursing,Xingtai Medical College,Hebei Province,Xingtai 054002,China;Department of Stomatology,Xingtai Medical College,Hebei Province,Xingtai 054002,China;Department of Neurobiology,School of Basic Medical Sciences,Heibei Medical University,Shijiazhuang 050017,China)

机构地区：[1]邢台医学高等专科学校公共教学部,河北邢台054002 [2]邢台医学高等专科学校基础医学部,河北邢台054002 [3]邢台医学高等专科学校护理系,河北邢台054002 [4]邢台医学高等专科学校口腔医学系,河北邢台054002 [5]河北医科大学基础医学院神经生物室,河北石家庄050017

出　　处：《河北医科大学学报》2019年第3期258-262,共5页Journal of Hebei Medical University

摘　　要：目的建立一种简便、易行的ICR小鼠神经干细胞(neural stem cells,NSCs)体外分离培养方法。方法分离胚胎ICR小鼠的脑组织,用无血清培养基悬浮培养;倒置显微镜观察细胞形态;免疫细胞化学染色检测特异性标记物神经上皮干细胞蛋白(巢蛋白,nestin)、胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)、神经元特异性烯醇化酶(neuronal specific enolase,NSE);CCK8法绘制生长曲线。结果从ICR小鼠胚脑组织分离的细胞,在无血清培养液中以神经球形式扩增,并可经消化后连续传代。生长曲线显示细胞倍增时间为2d。神经球内nestin阳性细胞率为(72.0±5.5)%。诱导分化后可观察到2种形态的细胞:一种具有长而粗的突起,交织成网,且GFAP阳性;另一种为胞体呈多边形、具有数个细长突起,NSE阳性。结论本方法培养的NSCs纯度高、增殖快,具有多向分化潜能,值得推广应用。Objective To establish a simple method for isolation and culture of mouse neural stem cells(NSCs)from ICR mice in vitro.Methods The brain tissues of ICR mice were isolated and cultured in serum-free medium.The morphology of cultured cells was observed with an inverted microscope.The specific markers,nestin,glial fibrillary acidic protein(GFAP)and neuronal specific enolase(NSE)were detected by immunocytochemical staining to identify cells.The growth curve of the cells was plotted by CCK8 assay.Results The cells isolated from the embryonic brain tissue of ICR mice were amplified in the form of neurospheres in serum-free medium,and could be passaged continuously after digestion.The growth curve showed that the cell doubling time was 2 d.The percentage of nestin positive cells was(72.0±5.5)%in neurospheres.After induced differentiation,two types of cells were observed:one has long and thick projection interlaced into a net,and is GFAP positive;the other is polygonal,with a number of slender projections,and is NSE positive.Conclusion The NSCs cultured by this method has high purity,proliferate rapidly,and have the potential of multi-differentiation.Our method is worth popularizing and applying.

关 键 词：神经干细胞 细胞培养技术 小鼠 

分 类 号：R394.26[医药卫生—医学遗传学]