犬骨髓间充质干细胞向血管内皮细胞定向分化的研究  

Experimental studies on differentiation of canine bone marrow mesenchymal stem cells into vascular endothelioid cells in vitro

作　　者：李春民[1] 董建德[2] 谷涌泉[3] 陈晓波[1] 卞策[1] 汪忠镐[3] 

机构地区：[1]首都医科大学附属复兴医院血管外科,北京100038 [2]北京电力总医院胸心血管外科,北京100073 [3]首都医科大学宣武医院血管外科,北京100053

出　　处：《中国现代普通外科进展》2010年第10期757-760,共4页Chinese Journal of Current Advances in General Surgery

基　　金：国家高技术研究发展计划(2006AA02A134);国家自然科学基金科学部主任基金(30640078)

摘　　要：目的:研究犬骨髓间充质干细胞(MSCs)在体外定向诱导,向血管内皮细胞(ECs)方向分化,探讨其作为组织工程血管种子细胞的可行性。方法:获取犬骨髓单个核细胞,体外分离出MSCs。将分离的MSCs接种于含EBM-2完全培养液的培养瓶,向内皮样细胞定向诱导分化;LG-DMEM完全培养液培养的MSCs作为对照组。对MSCs表型进行流式细胞仪鉴定;诱导的内皮样细胞进行CD31、Ⅷ因子免疫细胞化学鉴定和摄取Dil-Ac-LDL、结合FITC-UEA的双染色功能鉴定。结果:流式细胞术检测培养的原代MSCs表面标志CD44表达呈阳性,而CD34表达则呈阴性;诱导分化的内皮样细胞具有内皮细胞形态特征,与成体内皮细胞相似;诱导的内皮样细胞Ⅷ因子染色和CD31免疫荧光染色均呈明显阳性,对照组细胞为阴性;诱导的内皮样细胞Dil-Ac-LDL摄取实验和FITC-UEA结合实验,细胞呈阳性表现。结论:在体外用骨髓MSCs能够成功诱导分化为血管内皮样细胞,为组织工程血管内皮化提供了一种可行的种子细胞来源。Objective：To study the differenation of canine marrow mesenchymal stem cells（MSCs） into vascular endothelioid cells in vitro and to provide a new cells seed source for vascular tissue engineering.Methods： The MSCs were isolated from canine marrow mononuclear cells.The MSCs were seeded in EBM-2 differentiated into endothelioid cells and cultured in LG-DMEM as control group.The phenotype of MSCs was analyzed with flow cytometry.The endothelioid cells were tested by immunocytochemistry staining of vWF and CD31 and uptake of Dil-ac-LDL and binding of FITC-UEA.Results：The primary MSCs expression of CD44 but negative expression of CD34 as measured by flow cytometry.The morphologic characteristics of endothelioid cells were similar as the endothelial cells of adult.The endothelioid cells immunofluorescent staining of vWF and CD31 were positive and the cells of control group were negative.The endothelioid cells were Dil-ac-LDL and FITC-UEA double positive.Conclusion：The bone marrow MSCs can differentiate into vascular endothelioid cells after induced in vitro,which can provide sources of seed cells for vascular tissue engineering.

关 键 词：血管 组织工程 间质干细胞 内皮细胞 细胞分化 狗 

分 类 号：R318.08[医药卫生—生物医学工程] R654[医药卫生—基础医学]