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【目的】探讨体外诱导骨髓单个核细胞分化成为内皮祖细胞,为小径聚氨酯人工血管内皮化提供合适的种子细胞的可行性。【方法】收集健康成人骨髓单个核细胞,置于纤维连接蛋白预衬的DMEM培养基中,用血管内皮生长因子(vascular endothelial growth factor,VEGF)和碱性成纤维细胞生长因子(basic- fibroblast growth factor,bFGF)加以诱导,通过荧光显微镜和免疫组化分析等方法观察和鉴定诱导后的细胞。将诱导分化的内皮祖细胞种植到聚氨酯小径人工血管表面,用扫描电镜观察。【结果】在VEGF、bFGF等诱导因子存在的条件下,骨髓单个核细胞分化成为内皮祖细胞,倒置荧光显微镜下呈典形的“纺锤样”梭形细胞,单层细胞贴壁生长融合时呈铺路石样排列,免疫组化示VWF和CD34抗体染色阳性。扫描电镜下,未种植细胞的聚氨酯小径人工血管表面呈典型的多孔蜂窝状结构,孔径大小比较适合内皮祖细胞爬行。种植细胞后,聚氨酯小径人工血管表面有大量的内皮祖细胞黏附、爬行及铺展生长,有时可见内皮祖细胞长入蜂窝状孔径内。【结论】体外诱导骨髓单个核细胞分化为内皮祖细胞可作为小径聚氨酯人工血管内皮化的种子细胞。
【Objective】 To investigate the feasibility of differentiating bone marrow mononuclear cells into endothelial progenitor cells in vitro and providing appropriate seed cells for small-diameter polyurethane vascular endothelialization. 【Methods】 Bone marrow mononuclear cells of healthy adults were collected and placed in fibronectin-lined DMEM medium. VEGF, basic-fibroblast growth factor (VEGF) and basic fibroblast growth factor factor, bFGF) were induced to observe and identify the induced cells by fluorescence microscopy and immunohistochemical analysis. Differentiated endothelial progenitor cells were seeded on the surface of urethane-coated artificial blood vessels and observed by scanning electron microscopy. 【Results】 Under the conditions of inducing factors such as VEGF and bFGF, the differentiation of bone marrow mononuclear cells into endothelial progenitor cells was typical “spindle-like” spindle cells under inverted fluorescence microscope. Paving stone-like arrangement, immunohistochemistry showed VWF and CD34 antibody staining. Under the scanning electron microscope (SEM), the uninoculated cells showed a typical porous honeycomb structure on the surface of the vascular prosthesis. The pore size was suitable for the crawling of endothelial progenitor cells. After planting cells, a large number of endothelial progenitor cells adhere to, crawl and spread on the surface of the polyurethane small-diameter vascular prosthesis, and sometimes endothelial progenitor cells grow into honeycomb-shaped pores. 【Conclusion】 Differentiation of bone marrow mononuclear cells into endothelial progenitor cells in vitro can be used as seed cells of small-diameter polyurethane vascular endothelial cells.