胚脑来源与骨髓来源神经干细胞的比较:应用原子力显微镜的观察  

Comparison of neural stem cells derived from bone marrow and embryonic brain: observation under atomic force microscope

作　　者：周虎田[1] 徐如祥[1] 姜晓丹[1] 蔡颖谦[1] 郁毅刚[1] 邹雨汐[1] 杜谋选[1] 

机构地区：[1]南方医科大学珠江医院神经外科,广东省广州市510282

出　　处：《中国临床康复》2005年第5期45-47,i001,共4页Chinese Journal of Clinical Rehabilitation

基　　金：国家自然科学基金(30270491);广东省重大科技项目基金犤粤科基办(2004)08;粤财企(2001)367;(2003)209犦资助~~

摘　　要：目的:应用原子力显微镜(atomicforcemicroscopy,AFM)观测胚鼠前脑来源的神经干细胞及成鼠骨髓诱导来源的神经干细胞表面的形貌结构,并比较两者的异同。方法:实验于2003-09/2004-09在南方医科大学珠江医院全军神经医学研究所完成。分别取怀孕10.5～14.5d胚鼠行前脑来源神经干细胞和成鼠骨髓来源神经干细胞培养,神经干细胞培养基、碱性成纤维细胞生长因子(bFGF,20μg/L)维甲酸(300μg/L)持续诱导培养10d,以平板克隆法测定两种来源干细胞的克隆形成率,光学显微镜和原子力显微镜分别进行观测。结果:光学显微镜下两种来源的神经干细胞均表现为大圆形细胞,直径6～12μm,细胞形貌相似;但神经来源神经干细胞显示更强的增殖能力,其细胞克隆形成率显著大于骨髓来源神经干细胞(χ2=4.444,P<0.05);原子力显微镜下可观测到神经来源干细胞表面有较多颗粒样隆起,其表面粗糙,平均粗糙度为(18.5±2.0)nm,均方根粗糙度为(20.7±2.5)nm,而骨髓来源神经干细胞表面相对光滑,其表面平均粗糙度为(10.2±1.2)nm,均方根粗糙度为(12.9±1.3)nm,两者经t检验统计分析差异有非常显著性意义(P<0.01)。结论:神经来源神经干细胞较骨髓来源神经干细胞表面有更多颗粒样隆起,表面更粗糙,可能与其有更强的增殖能力有关。AIM:To observe the surface morphology of neural stem cells(NSCs) derived from bone marrow of adult rats and embryonic rat brain under atomic force microscopy(AFM) and compare the difference. METHODS:The experiment was finished in Institute of Neurology of Chinese PLA,Zhujiang Hospital of Nanfang Medical University from September 2003 to September 2004.The NSCs derived from embryonic rat forebrain at enceinte 10.5-14.5 days and NSCs from the bone marrow of adults rats were experimentally cultured respectively.NSCs culture medium,basic fibroblast growth factor(bFGF,20 μg/L1),and retinoic acid(300 μg/L) were continuously used to culture and induce in vitro for 10 days.The rate of cell clone was measured by plate cultivation cloning technique. The cell morphology and structure were observed with light microscopy and atomic force microscope. RESULTS:The form of NSCs derived from both of the different tissues was alike under light microscope.Both of them showed big and round morphology, and their diameters were 6-12 μm.But NSCs derived from embryonic brain showed more strong proliferation ability,and were much more reliable to formatting clone than those from bone marrow(χ2=4.444,P< 0.05).The NSCs derived from embryonic brain showed rougher surface structure and more granules on cell surface than those derived from bone marrow under AFM.The term of average roughness and mean square roughness of the neural stem cells derived from embryonic brain were(18.5±2.0) nm and(20.7±2.5) nm,but the ones of the neural stem cells derived from bone marrow were(10.2±1.2) nm and(12.9±1.3) nm respectively.Both of them showed significant difference in t test(P< 0.01). CONCLUSION:The NSCs derived from embryonic rat brain have rougher surface structure and more granules than those derived from bone marrow.It may be related to the ability of the cell proliferation.

关 键 词：干细胞 神经系统/细胞学 显微镜检查 原子力 

分 类 号：R743[医药卫生—神经病学与精神病学]