异种骨对神经干细胞诱导分化的作用  

Effect of xenogeneic bone on induced differentiation of neural stem cells

作　　者：仇士东[1] 吴世福[1] 孙晓霞[1] 张磊[2] 李娜[1] 侯丽[1] 

机构地区：[1]山东省医疗器械产品质量检验中心山东省医疗器械生物学评价重点实验室,济南250101 [2]山东省医疗器械产品质量检验中心包材室,济南250101

出　　处：《广东医学》2015年第17期2644-2648,共5页Guangdong Medical Journal

基　　金：再生型医用植入器械国家工程实验室PI项目(编号:2012NELRMD007);"十二五"国家科技支撑计划项目(编号:2012BAI22B01)

摘　　要：目的探讨异种骨对大鼠神经干细胞(NSCs)诱导分化的作用。方法从15 d胎龄大鼠的大脑皮质分离获得NSCs后,以神经球悬浮培养法,采用DMEM/F12为基础培养基,加入2%B27、20μg/L EGF和20μg/L b FGF等进行培养。传代3次后,采用巢蛋白染色,免疫荧光细胞化学法鉴定NSCs。然后,加入浓度分别为0.025、0.05、0.1、0.2 g/m L的异种骨浸提液与NSCs进行共培养。结果传代后的细胞表现出NSCs的特性。与0.2 g/m L异种骨浸提液共培养的NSCs微管相关蛋白、胶原纤维酸性蛋白和O4均呈阳性表达,即异种骨浸提液可显著诱导NSCs分化为神经元、星形胶质细胞和少突胶质细胞;其中异种骨浸提液浓度为0.1 g/m L时,诱导NSCs分化为少突胶质细胞的比例最高;浓度为0.05 g/m L时,分化为星形胶质细胞的比例最高。结论异种骨具有诱导NSCs分化为神经元及胶质细胞的作用,且浓度不同,诱导分化的细胞比例成分亦不同。临床应用中,可结合一定的生长因子,诱导其分化方向,从而有利于修复神经系统损伤。Objective To investigate the effect of xenogeneic bone on induced differentiation of neural stem cells（ NSCs）. Methods NSCs were isolated from cerebral cortex of the frontal lobe in 15 d rat embryos and were cultured using the neurosphere culture system. The basic culture medium was DMEM / F12,added with 2% B27,20 μg / L EGF,20μg / L b FGF and others. After 3 passages,NSCs were stained by Nestin and identified by immunocytochemistry. They were then co- cultured with xenogeneic bone extracts at concentrations of 0. 025,0. 05,0. 1,0. 2 g / m L. Results Cells after passage exhibited characteristics of NSCs. Positive expression of microtubule- associated protein 2,glial fibrillary acidic protein and O4 were detected in NSCs co- cultured with 0. 2 g / m L xenogeneic bone extracts. That is to say,xenogeneic bone extracts could induce NSCs differentiate into neurons,astrocytes and oligodendrocyte. When the concentration of xenogeneic bone extracts was 0. 1 g / m L,the proportion of NSCs that differentiate into oligodendrocytes was the most. When the concentration was 0. 05 g / m L,the proportion of NSCs that differentiate into astrocytes was the most. Conclusion Xenogeneic bone can induce NSCs to differentiate into neurons and glial cells. The proportion of differentiated cells was different at various concentrations of xenogeneic bone. Therefore,in its clinical application,the xenogeneic bone combined with certain growth factors to induce NSC differentiation can help repair the damage of the nervous system.

关 键 词：神经干细胞 异种骨 分化 神经元 

分 类 号：R733.71[医药卫生—肿瘤]