兔骨髓基质干细胞向血管内皮样细胞的诱导分化  

Differentiation of Rabbits' Bone Marrow Mesenchymal Stem Cells into Vascular Endothelial Like Cells

作　　者：翁雁鸣 王之发 居兆慧 刘斌[3] 刘彦普 

机构地区：[1]第四军医大学口腔医院颌面外科,陕西西安710032 [2]第四军医大学口腔医院儿童口腔科,陕西西安710032 [3]第四军医大学口腔医院口腔生物学教研室,陕西西安710032

出　　处：《现代生物医学进展》2013年第12期2225-2229,共5页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(81070820;31170942);陕西省"13115"科技创新工程重大科技专项项目(2009ZDKG-74)

摘　　要：目的:研究血管内皮细胞生长因子(VEGF)联合碱性成纤维细胞生长因子(bFGF)促进兔骨髓基质干细胞向血管内皮样细胞的定向诱导分化,为血管化组织工程骨研究提供实验基础。方法:采集2周龄兔后肢长骨骨髓,用全骨骨髓贴壁法进行原代培养,将获得的第2代骨髓基质干细胞以1×105/mL密度接种于内皮细胞条件培养基(含10μg/L VEGF,10μg/L bFGF,10%胎牛血清的DMEM/F12培养液)进行体外诱导培养,对诱导2周的细胞进行细胞形态观察和表型、功能鉴定。结果:经血管内皮细胞条件培养基诱导2周后的细胞呈扁平形,多边形,表达血管内皮细胞特异性标志CD31、VWF因子,细胞具有吞噬DiI-Ac-LDL和摄取FITC-UEA-1的功能,诱导的细胞可在BD基质胶内形成管腔样结构。结论:血管内皮细胞生长因子联合碱性成纤维细胞生长因子可以成功诱导兔骨髓基质干细胞为血管内皮样细胞,有希望作为组织工程骨的血管化的种子细胞。Objective： To obtain and characterize vascular endothelial like cells derived from rabbits＇ bone marrow mesenchymal stem cells （BMSCs） in vitro, and to provide a kind of new cells seed source for vascularized bone tissue engineering. Methods： BMSCs were isolated from rabbit bone marrow. The second generated BMSCs were cultured in the DMEM medium containing 10μg/L vascular endothelial growth factor （VEGF）, 10μg/L basic fibrioblast growth factor （bFGF） and 10 % fetal bovine serum for 14 days. The endothelial like cells were identified by morphological traits, immunofluorescent staining and functional tests. Results： Cells induced from BMSCs exhibited as spindle or polygonal shaped cells. Indirect immunofluorescent staining date revealed the highly expression of CD31 and yon Willebrand factor on the surface of the induced cells. The cells can incorporate DiI-ac-LDL and uptake FITC-UEA-1 lectin. When grown on BD MatrigelTM, induced cells formed tube-like structures. Conclusion： The combination of VEGF with bFGF can successfully induce differentiation of rabbit bone marrow mesenchymal stem cells into vascular endothelial like cells, which may be as the sources of seed cells for vascular bone tissue engineering.

关 键 词：骨髓基质干细胞 血管内皮样细胞 血管内皮生长因子 碱性成纤维细胞生长因子 

分 类 号：Q953[生物学—动物学] Q75