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1、高表达Ang1基因的间充质干细胞的建立及其肺内转化研究徐金富瞿介明李惠萍等同济大学附属上海市肺科医院复旦大学附属华东医院Highpromiseforangiopoietin-1(Ang1)inlunginjuryinterventionAng1decreasedinLPSinducedlunginjuryanti-inflammation:inhibitsendothelialpermeability,neutrophiladherenceandIL-8productionAnti-apoptosisroletoendothelialcellsTh
2、urstonG,RudgeJS,IoffeE,etal.Angiopoietin-1protectstheadultvasculatureagainstplasmaleakage.NatMed.2000,6:460-463. DalyC,WongV,BurovaE,etal.Angiopoietin-1modulatesendothelialcellfunctionandgeneexpressionviathetranscriptionfactorFKHR(FOXO1).GeneDev.2004,18(9):1060-1071.PizurkiL,Z
3、houZ,GlynosK,etal.Angiopoietin-1inhibitsendothelialpermeability,neutrophiladherenceandIL-8production.BritJPharmacol.2003,139(2):329-336.Mesenchymalstemcells(MSC)NoethicaldifficultiesharvestMSCfromtheindividualtobetreatedNorejectionproblemsSelfmigratetowheretheinjuryislocatedRo
4、jasM,XuJ,WoodsCR,etal.Bonemarrow-derivedmesenchymalstemcellsinrepairoftheinjuredlung.AmJRespirCellMolBiol2005;33:145–152 RobertJD，AnnemarieBM．mesenchymalstemcells：biologyandpotentialclinicaluses.ExperimentalHematology,2000,28(8):87Ourhypothesis+Ang1MSCsByLentiviralvectorMSCstr
5、ansducedwithAng1MiceLPSinducedlunginjuryInvivointerventionMethodsandresultspCR2.1-Ang1:Prof.FedericoBussolinoinInstituteforcancerresearchandtreatment,ItalySequencingresults:100%coincidewithAng1cDNAsequenceElectrophoresisaftercuttingbyrestrictionenzymepLVpCMV-dR8.74pMD2GInvitro
6、Lentivirus-mediatedAng1/eGFPtransferintoMSCsIsolation,purificationandexpansionofMSCsBonemarrowwasharvestedfrom6weeksoldC57BL/6byflushingthefemursandtibiasSeparateMSCsfrombonemarrowbydensitygradientcentrifugationinFicoll(Pharmacia)CulturedandpassagedwithPeister’smethodPeisterA,
7、MelladJA,LarsonBL,HallBM,GibsonLF,ProckopDJ.Adultstemcellsfrombonemarrow(MSCs)isolatedfromdifferentstrainsofinbredmicevaryinsurfaceepitopes,ratesofproliferation,anddifferentiationpotential.Blood2004;103:1662-1668.MSCstransducedwithLenti-Ang1/eGFP1×106MSCsfrompassage4wereseeded
8、intoa12-wellplateMultiplicityofinfection(MOI):10，20，40，80Merg
