【摘要】 目的:分析人骨髓间充质干细胞(hMSCs)和脐静脉内皮细胞(hUVECs)的基因表达差异,探讨体外基因转染诱导内皮分化的可行性以及作为血管组织工程种子细胞来源的应用前景。方法:分别从人骨髓和脐静脉分离间充质干细胞(hMSCs)和内皮细胞(hUVECs),扩增培养后进行流式细胞仪、免疫细胞化学,免疫荧光鉴定和超微结构观察。通过BiostarH-40S表达谱芯片分析,选择两者的差异表达基因,导入hMSCs,经RT-PCR、ELISA鉴定该基因的转染和表达,并分析hMSCs的内皮分化程度。结果:hMSCs表达内皮细胞的多种特异性mRNA,经VEGFl65基因瞬时转染后RT-PCR有明显条带,ELISA定量检测VEGF165蛋白表达为(707.9±11.3)ng/L,同时CD44表达明显下调38.80%,CD31则明显上调达56.82%,FI-1,FVⅢAg和CD34的表达也有不同程度升高。结论:hMSCs具有内皮分化潜能,体外基因转染诱导hMSCs产生功能性内皮细胞和组织工程化血管具有广阔前景。更多还原

【Abstract】 Aim: To analyze gene expression difference between human mesenchymal stem cells and umbilical vein endothelial cells, to discuss the feasibility of inducing hMSCs to differentiate intoendothelial cells through in vitro gene transfection as well as the use and prospective as a seedingcell source of vascular tissue engineering. Methods: hMSCs and hUVECs were isolated,expanded in culture, and characterized by flow-cytometry, immunocytochemistry,immunofluorescence and transmission electron microscopy (TEM). Di fferential analysis of geneexpression profiles between them was performed by Biostar H-40 cDNA microarrays. The properties of VEGF 165 transfected were also detected with RT-PCR, ELISA et al. Results: Nearly 86% genes were coexpressed in both cells and hMSCs expressed typical endothelial antigenmarker of EC. After VEGF165 transfection, hMSCs (or committed hMSCs ) were positive forCD31. To different extent, the expression of CD44 was down regulated and CD34, FVⅢAg, Flt-1up regulated. Conclusion:Generation of functional EC or tissue engineered blood vessels fromhuman MSCs is feasible utilizing an in vitro environment gene [LM]transfection.更多还原