骨髓间充质干细胞诱导分化为血管内皮样细胞的实验的研究

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1、摘要碱性成纤维细胞生长因子；体外培养；组织工程IIIAbstractABSTRACTObjectiveTostudythefeasibilityforinducingtodifferentiatemesenchymalstemcells(MSCs)intovascularendothelial．1ikecellswhichwereisolatedandpurifiedinvitro，toobservetheinducedvascularendothelial-likecellswhethertoparticipateintowoundsurfaceangiopoiesisth

2、roughanimalexperiment．Methods111epurifiedbonemarrowmesenchymalstemcellswith2mlwereobtainedfrompatientswithouttheseverediseasesinclinicalexaminationafterFicolldensitygradientcentrifugationandmonolayerculturedofthecollectedbonemarrowaspiration，thesecellswereculturedandpassagedinvitro．Theywere

3、observed、析tllinvertedphasecontrastmicroscopeformorphologicalobservationandwereidentifiedbytheflowcytometry、scanningelectronmicroscope、transmissionelectronmicroscope、immunocytochemistrymethods．W．eusedL4(2’)orthogonalexperimentaldesigntoboltinductionfactorsvascularendothelialgrowthfactor(VEGF

4、)andbasicfibroblastgrowthfactor(bFGF)，andincubatedMSCsintovascularendothelial-likecellsaccordingtotheboltingresults．Themorphologicalobservationofinducedcellswereobserved、析thinvertedphasecontrastmicroscope，theinducedcellswereidentifiedbyscanningelectronmicroscope、transmissionelectronmicrosco

5、pemethods、theimmunocytochemistrymethods．TissueengineeringdermissurrogateWasconstructed、析tllcollagenproteinspongeframeandvascularendothelial-likecellswhichweremarkedby5-bromodeoxyuridine．耶1efull-thicknessskindefectedwoundsurfaceinrabbitbackwerecuredbythetissueengineeringdermissurrogate．After

6、14days，weusedwoundsurfacetissueskintodotheimmunohistochemistrystaining．ResultsneappearanceofMSCswhichwereobtainedbyFicolldensitygradientcentrifugationandmonolayerculturedisfusiform，theshortpachymicrovilliofcellsurfaceandcellmatrixweresawbyscanningelectronmicroscope．Twoarchitectureswerecorre

7、spondedtothedifferentcell’Sfunctionstatesbytransmissionelectronmicroscope，flowcytometryshowedthat85．35percentofcellswerefoundatGo、GIphase．ExpressionofCD29、CD44werepositiveonMSCsIVAbstractsurfaceandCD34、CD45、Ⅷfactorwerenegative．ComparetheⅧfactorpositiveex