【摘要】 目的:探讨人骨髓间充质干细胞(Bone Mesenchymal Stem Cells,BMSC)分化形成的神经样细胞的形态及结构特征。方法:采用密度梯度离心法和贴壁培养的方法分离纯化人BMSC,流式细胞仪检测CD34、CD45、CD44等细胞表面抗原在BMSC上的表达。联合应用碱性成纤维细胞生长因子(basic fibroblast growthfactor,bFGF)、表皮生长因子(epidermal growth factor,EGF)、二甲基亚枫(di methyl sulphoxide,DMSO)和丁羟基茴香醚(butylhydroxy anisol,BHA)诱导人BMSC向神经细胞分化。光学显微镜下观察细胞的形态变化;甲苯胺蓝染色法对分化后细胞进行尼氏体染色;透射电子显微镜观察分化后细胞的超微结构特点。结果:分离纯化的人BMSC经流式细胞仪检测显示间充质干细胞相对特异性抗原CD44表达阳性,造血干细胞特异性抗原CD34、CD45表达阴性;加入诱导培养基培养2周后,细胞胞体变圆,折光性增强,有突起形成;甲苯胺蓝染色法显示分化后的细胞尼氏体染色阳性;透射电镜下细胞胞浆中有大量密集的呈扁平囊状的粗面内质网及其间的一些游离核糖体和多聚核糖体所组成的尼氏体的超微结构,并有神经微丝的形成。结论:EGF/bFGF/DMSO/BHA诱导人BMSC分化形成的神经样细胞能够表达成熟神经细胞典型的形态及超微结构特征。更多还原

【Abstract】 Objective: To investigated the shape and ultrastructure of the neuron-like cells derived from human BMSC,thus to explore whether the neuron-like cells derived from human BMSC possessed the shape and ultrastructure of mature neurons or not.Methods: Human BMSC were separated from adult bone marrow with density gradient centrifugation and adherent culture,induced and differentiated into neuron-like cells with EGF/ bFGF/DMSO/BHA.The expression of CD34,CD44,CD45 and other antigen on human BMSC were detected by flow cytometer(FCM),Nissl body by toluidine blue staining,ultrastructure of the neuron-like cells by electron microscope.Results: Flow cytometry displayed cells were positive for CD44,showing a typical mesenchymal like immunophenotype,on the other hand,they were negative for CD45 and CD34 markers associated with hematopoietic cells.After induced 2w by EGF/ bFGF/DMSO/BHA,almost bulk of cells had morphological features typical of neurons,such as spherical shape and extending processes.Analysis confirmed that toluidine blue staining for Nissl body had been induced by EGF/ bFGF/DMSO/BHA.Using electron microscope,we could also see lots of flat,vesicular rough endoplasmic reticulum(RER),some free ribosome and neurofilament in the cytoplasm.Conclusion: Neuron-like cells derived from human BMSC can express not only the typical shape of mature neurons but also the ultrastructure of mature neurons.更多还原