细胞间直接接触对骨髓间充质干细胞分化为血管内皮细胞的作用.docVIP

PAGE 细胞间直接接触对骨髓间充质干细胞分化为血管内皮细胞的作用 彭锦，迟路湘 （400038重庆 ，第三军医大学西南医院心血管内科） [摘要] 目的：探讨骨髓间充质干细胞(mesenchymal stem cells, MSCs)与人脐静脉内皮细胞(human umbilical vein endothelial cells, HUVECs)直接接触共培养诱导MSCs向血管内皮细胞分化的作用。方法：将MSCs和HUVECs按一定比例混合直接共培养，48 h后用ELISA检测其共培养细胞培养液（实验组）及MSCs与HUVECs各自单独培养48h后再混合的培养液（对照组）中VEGF含量；5天后用免疫荧光细胞化学法观察和分析诱导后MSCs的胎肝激酶-1 ( fetal liver kinase-1, Flk-1) 和血管性血友病因子( von Willebrand factor, vWF)的表达以及其Dil-ac-LDL吞噬能力；并用透射电镜观察MSCs超微结构改变。结果：VEGF在实验组中的含量显著高于对照组；直接接触共培养诱导MSCs表达Flk-1蛋白,且部分Flk-1阳性MSCs同时表达vWF蛋白；部分MSCs具有吞噬Dil-ac-LDL功能；电镜下见MSCs出现分化特征，核质比缩小，细胞器较为丰富，并可见细胞融合现象。结论：HUVECs可以通过细胞间直接接触诱导共培养的MSCs开始向内皮分化；其机制可能与细胞间直接接触促进细胞分泌VEGF以及细胞融合密切相关。 [关键词] 间充质干细胞；脐静脉内皮细胞；共培养；分化；VEGF；超微结构 Effect of cell-cell direct contact to mesenchymal stem cells differentiate into vascular endothelial cells Peng Jin, Chi Luxiang (the Department of Cardiovasology, Southwest Hospital, Third Military Medical University,chongqing,China 400038) [Abstract] Objective 　To investigate the effect of cell-cell direct contact to mesenchymal stem cells(MSCs) differentiate into vascular endothelial cells(VECs) when MSCs co-cultured with human umbilical vein endothelial cells（HUVECs）in vitro. Methods MSCs and HUVECs were cultured and identified，and then co-culture. The VEGF levels of experimental group (culture media of co-cultured for 48 hours) and control group (MSCs and HUVECs’s mixed culture media after them were cultured alone for 48 hours) were confirmed using ELISA assays. Co-cultured for 5 days, The MSCs’s expression of fetal liver kinase-1(Flk-1) and von Willebrand factor (vWF) and ingestible ability of Dil-ac-LDL was analyzed by immunofluorescence staining,and the changes of ultrastructure of co-cultured cells were observed. 　Results 　 The VEGF levels of experimental group was significantly higher than control group； In direct contact co-culture system，some MSCs were induced to express Flk-1 protein, and part of the Flk-1 positive MSCs also expressed vWF protein ；some MSCs could phagocy