不同类型星形胶质细胞对神经干细胞定向分化的影响  

Effects of Different Astrocytes on Differentiation of Neural Stem Cells

作　　者：刘媛[1] 龙在云[1] 曾琳[1] 李民[1] 伍亚民[1] 王正国[1] 

机构地区：[1]第三军医大学大坪医院野战外科研究所三室,创伤,烧伤与复合伤国家重点实验室,重庆市400042

出　　处：《中国康复理论与实践》2007年第11期1026-1028,F0003,共4页Chinese Journal of Rehabilitation Theory and Practice

基　　金：重庆市院士基金(合同号:7671)资助;国家自然科学基金(30171068)资助

摘　　要：目的观察不同类型星形胶质细胞作为饲养层细胞对神经干细胞定向分化为神经元的影响。方法通过无血清培养的方法,将原代培养所获得的神经干细胞克隆;经纯化和标记后,分别接种在以原浆型和纤维型星形胶质细胞作为饲养细胞的培养皿中并加入NeuralBasal培养基,10d后行NF-200免疫荧光染色,在显微镜下随机选取20个视野,记数神经元和标记神经干细胞的比例,并对分化的神经元细胞进行胆碱酯酶染色和Fura-3探针标记。结果在原浆型和纤维型星形胶质细胞培养条件下,神经干细胞分化为神经元的比例分别为72%与43%,分化细胞胆碱酯酶染色阳性。分化神经元在药物刺激下,细胞内可发生钙离子活动的变化。结论原浆型星形胶质细胞具有较好地促进神经干细胞向神经元,特别是向有生物学活性的运动神经元分化,可作为神经组织工程中研究中一种新的种子细胞。Objective To observe the differentiation of neural stem cells （NSC） into neurons under different astrocytes feeding layers. Methods The NSC purified from primary cultured clones and labeled by DAPI in serum-free media were plated in different feeding layers： cyto-plasma astrocytes and fibrous astrocytes respectively with Neural Basal （NB） media. After 10 d, immunohisto- chemistry with antibody NF-200 was taken to calculate the percents of neurons by 20 fields of vision chosen randomly. The differentiated neurons were stained with Ache and labeled with Fura-3AM. Results The ratios of differentiated neurons in cyto-plasma and fibrous astrocytes were 72% and 43% respectively. In most differentiated neurons the AchE staining was positive and had the activity of Ca2＋ stimulated by medicine. Conclusion The cyto-plasrna astrocytes can induced NSC differentiated into neurons, especially into active motor neurons, which can be chosen for a new seeding cells in the nervous tissue engineering.

关 键 词：星形胶质细胞 原浆型 纤维型 神经干细胞 神经元 分化 

分 类 号：R318.5[医药卫生—生物医学工程]