人骨髓间充质干细胞体外分离培养及向血管内皮样细胞分化的研究  

Isolating culture of human bone marrow-derived mesenchymal stem cells and the differentiation into blood vessel endothelial-like cells in vitro

作　　者：冯海燕[1] 刘瑞风[2] 张开明[2] 

机构地区：[1]山西医科大学第二医院,太原030000 [2]山西省太原市中心医院皮肤科,太原030000

出　　处：《中国免疫学杂志》2010年第1期70-74,78,共6页Chinese Journal of Immunology

基　　金：国家自然科学基金资助项目(No.30771940)

摘　　要：目的:研究人骨髓间充质干细胞(hMSCs)的体外分离培养方法及在特定微环境下分化为血管内皮样细胞的潜能,可能为银屑病研究提供实验基础。方法:密度梯度离心法结合贴壁法分离人骨髓间充质干细胞(hMSCs),体外扩增培养并进行鉴定。加入血管内皮细胞生长因子(VEGF)、碱性成纤维细胞生长因子(bFGF)定向诱导hMSCs向血管内皮细胞分化,流式细胞仪进行细胞表型鉴定,Dil-ac-LDL摄取实验鉴定血管内皮细胞功能。结果:经流式细胞仪测定分离培养的hMSCsCD71、CD44阳性表达,CD54、CD106、CD45弱阳性表达,CD34、CD31、VWF、KDR、HLA-DR阴性表达;经bFGF、VEGF诱导后的hMSCs可见类似内皮细胞样改变,经流式细胞术检测,与对照组相比内皮细胞特异表面标志CD34、CD31、VWF、KDR表达转为阳性(P<0.01),而HLA-DR、CD54、CD106、CD45表达明显上调(P<0.01);诱导分化后的内皮样细胞具有摄取Dil-ac-LDL的能力。结论:密度梯度离心法结合贴壁法可获得较纯的MSCs;在bFGF、VEGF诱导作用下,hMSCs具有向内皮细胞分化潜能,产生功能性内皮细胞。Objective： To study of isolating culture and differentiation of human bone marrow-derived mesenchymal stem cells into blood vessel endothelial-like cells in a specialized micm-envimmnent in vitro, so as to provide an experimental foundation for psoriasis. Methods：The hMSCs were isolated by density gradient centrifugation, amplificated and identitlcated in vitro. Vascular endothelial growth factor （VEGF） and basic fibroblast growth factor （bFGF） within endothelial cell growth medium （DMEM） were used to induce hMSCs differentiation into vascular endothelial-like cells. The induced hMSCs were detected by flow cytometry to find whether they had endothelial cell phenotypes. The Dil-ac-LDL ingestion assay was used to appraises the blood vessel endothelial-like cell function. Results： In call morphology, the induced hMSCs tranformed into endothelial-like calls. These cells expressed specific surface markers of .vascular endothelial-like cells such as CD34, CD106, HLA-DR, CD54, VWF, CD31, KDR and CD45 comparing to those in the control group（ P 〈 0.01 ）. The induced endothelial-like cells had the ability of ingesting Dil-ac-LDL. Conclusion： Combination of Density gradient centrifugation and adherent methods can obtain pure MSCs.hMSCs can obtain endothelial cell phenotypes after indueed by VEGF and bFGF in vitro. Human hMSCs have potential to differentiate into vascular endothelial-like cells. The induced endothelial-like cells have completely mature endothelial cell functional properties.

关 键 词：骨髓间充质干细胞 血管内皮细胞 血管内皮细胞生长因子 碱性成纤维细胞生长因子 

分 类 号：R758.63[医药卫生—皮肤病学与性病学]