红细胞生成素激活的星形胶质细胞条件培养液对神经干细胞促分化作用及对分化后细胞的保护作用  

Effect of EPO-activated astrocyte conditioned medium on differention of neural stem cells and its protective effect on differentiated neural stem cells after injury in vitro

作　　者：张静[1] 沈慧[1] 夏春林[1] 

机构地区：[1]苏州大学细胞神经生物学研究室及衰老与神经疾病实验室

出　　处：《解剖学报》2009年第3期374-378,共5页Acta Anatomica Sinica

基　　金：国家自然科学基金资助项目(30772242);江苏省高校自然科学研究计划资助项目(05KJB310121)

摘　　要：目的观察红细胞生成素激活的星形胶质细胞条件培养液（EACM）对神经干细胞促分化作用及对分化后细胞的保护作用。方法原代培养神经干细胞和1型星形胶质细胞,收集红细胞生成素（EPO）刺激的星形胶质细胞上清液,用于神经干细胞分化实验的研究。对分化后的细胞进行免疫细胞化学染色,计算其分化为神经元的比率;同时利用FeSO4和H2O2制造细胞损伤模型,用EACM继续培养48h,然后检测细胞活性和存活细胞数。结果EACM组神经干细胞分化明显,神经元比率较星形胶质细胞上清组和对照组均有明显增加。分化后的细胞中加入H2O2后,继续用EACM培养的实验组吸光度（A）值和细胞存活百分比均较对照组高。结论红细胞生成素激活的星形胶质细胞条件培养液对神经干细胞有促其向神经元分化的作用,并对分化后的细胞有保护作用。Objective To investigate whether erythropoietin （EPO） activated astrocytes could release some bioactive factors which affect differentiation of neural stem cells and protect these differentiated cells after hydroxyl free radical induced injury. Methods Primarily cultured astrocytes were prepared and purified, and the astrocytes conditioned medium was collected and used to culture neural stem ceils. After 5 days in vitro the morphological changes of the neural stem cells were observed under a phase contrast microscope, NF-200 immunocytochemical staining was conducted to evaluate the differentiation of neural stem cells. In the mean time, FeSO4 and H2O2 were added to neural stem cells for 20 rain, and these cells were cultured for 48 hours with or without EACM, then the viability of cells was determined with MTT assays, and survived cells were counted as well. Results The expression of NF-200 in the neural stem cells showed strong positive whereas control group exhibited less morphological changes and NF-200 immunocytochemical staining was almost negative in corresponding period. As for EACM protective effects on differentiated neural stem cells after hydroxyl free-induced injuries generated by adding FeSO4 and H2O2, the absorbance value of the experimental group was 0.381 ± 0.027 while the data was only 0.27 ±0.013 in control group （P 〈 0.01 ）. Conclusion These results suggest that EPO activated astrocytes can secrete some factors to either promote differentiation of neural stem cells into neuron or protect differentiated neural stem cells under injury conditions in vitro.

关 键 词：神经干细胞 促红细胞生成素 分化 免疫组织化学 大鼠 

分 类 号：R322[医药卫生—人体解剖和组织胚胎学]