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

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1、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(mesenchymalstemcells,MSCs)与人脐静脉内皮细胞(humanumbilicalveinendothelialcells,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，48h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

2、细胞化学法观察和分析诱导后MSCs的胎肝激酶-1(fetalliverkinase-1,Flk-1)和血管性血友病因子(vonWillebrandfactor,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

3、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Effectofcell-celldirectcontacttomesenchymalstemcellsdifferentiateintovascularendothelialcellsPengJin,ChiLuxiang(theDepartmentofCardiovasology,SouthwestHospital,ThirdMilitaryMedicalUniversity,chongqing,China40

4、0038)[Abstract]Objective　Toinvestigatetheeffectofcell-celldirectcontacttomesenchymalstemcells(MSCs)differentiateintovascularendothelialcells(VECs)whenMSCsco-culturedwithhumanumbilicalveinendothelialcells（HUVECs）invitro.MethodsMSCsandHUVECswereculturedandidentified，andthenco-culture.The

5、VEGFlevelsofexperimentalgroup(culturemediaofco-culturedfor48hours)andcontrolgroup(MSCsandHUVECs’smixedculturemediaafterthemwereculturedalonefor48hours)wereconfirmedusingELISAassays.Co-culturedfor5days,TheMSCs’sexpressionoffetalliverkinase-1(Flk-1)andvonWillebrandfactor(vWF)andingestibl

6、eabilityofDil-ac-LDLwasanalyzedbyimmunofluorescencestaining,andthechangesofultrastructureofco-culturedcellswereobserved.　Results　TheVEGFlevelsofexperimentalgroupwassignificantlyhigherthancontrolgroup；Indirectcontactco-culturesystem，someMSCswereinducedtoexpressFlk-1protein,andpartoftheF

7、lk-1positiveMSCsalsoexpressedvWFprotein；someMSCscouldphagocytoseDil-ac-LDL;thereducedkaryoplasmicratioandabundantorganelleswereobservedintheco-culturedMSCsbytransmissionelectronmicroscopy,andcellfusioncanbeseen.Conclusion　HUVECsinduceMSCstodifferentiateintoendothelialcellsthroughdire