骨髓间充质干细胞定向分化为血管内皮样细胞的实验研究  

Study on the Differentiation of Human Mesenchymal Stem Cells into Vascular Endothelial-like Cells

作　　者：徐蓉[1] 徐金勇[2] 刘玮[1] 

机构地区：[1]安徽医学高等专科学校药学系,合肥230601 [2]安徽医科大学基础医学院人体解剖学教研室,合肥230032

出　　处：《生物医学工程学杂志》2014年第2期389-393,共5页Journal of Biomedical Engineering

基　　金：国家自然科学基金青年基金资助项目(81000589)

摘　　要：体外诱导人骨髓的间充质干细胞(MSCs)分化成为血管内皮样细胞(ELCs),探讨骨髓来源的MSCs作为组织工程血管种子细胞的可行性。通过密度梯度离心,分离人骨髓的单个核细胞,DMEM贴壁培养MSCs,多次传代贴壁培养纯化MSCs,包含血管内皮生长因子(VEGF)、人成纤维细胞生长因子(hFGF)、胰岛素样生长因子-1(IGF-1)和人表皮生长因子(hEGF)的EBM-2培养基诱导MSCs向ELCs分化,倒置显微镜观察ELCs的形态,流式细胞仪检测ELCs的血管内皮钙粘素(VE-cadherin)、CD133和CD31表达,免疫荧光检测ELCs的CD31和血管性假血友病因子(vWF)表达。结果显示,贴壁培养的MSCs呈长梭形,旋涡状生长,经过诱导细胞形态发生改变,细胞形态变圆,与血管内皮细胞(ECs)相似;流式细胞仪结果显示ELCs表达ECs的特异性标志物CD31和VE-cadherin,不表达CD133;免疫荧光结果也显示ELCs表达CD31和vWF。结果表明:MSCs具有定向诱导分化为ECs的潜能,MSCs来源的ELCs具有与ECs相似的细胞生物学特征,提示骨髓来源的MSCs可作为血管组织工程的种子细胞。To explore the feasibility of mesenchymal stem cells （MSCs） acting as seed cells in tissue engineering, we isolated human bone marrow MSCs and differentiated them into vascular endothelial-like cells （ELCs） in vitro. Bone marrow mononuclear cells （BMSCs） were isolated by the method of percoll density centrifugation, and seeded in Dul- becco Modified Eagle Medium supplemented with 10% fetal bovine serum. MSCs were purified through multiple ad- herent cultures, and differentiated into ELCs induced by endothelial cell growth medium-2 （EBM-2） medium contai- ning vascular endothelial growth factor （VEGF）, human fibroblast growth factor （hFGF）, insulin like growth fac- tors 1 （IGF-1）, and human epidermal growth factor （hEGF）. The relative biologic characteristics of ELCs including cell morphology and phenotype were studied by inverted microscope and flow cytometry. The induced cells were i- dentified by immunofluorescence with CD31 and Von Willebrand factor （vWF）. The results showed that the mor- phology of MSCs was long-spindle and vortex-like growth. After induction of differentiation, the cells were round, and similar to vascular endothelial cells （ECs）. Flow cytometric analysis revealed that ELCs expressed ECs specific surface markers of CD31 and vascular endothelial cadherin （VE-cadherin）, but not CD133. Immunofluorescence re- sults also confirmed that ELCs expressed CD31 and vWF. The results suggested that ELCs possed similar cell bio- logical characteristics with ECs. In one word, human MSCs derived from bone marrow have the potential to differen- tiate into ECs in vitro, and show clinical feasibility acting as ideal donor cells of vascular tissue engineering.

关 键 词：骨髓间充质干细胞 血管内皮样细胞 培养 分化 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]