肿瘤多细胞球_一种近似实体瘤的体外肿瘤模型

306201112JournalofChineseElectronMicroscopySocietyVol.30No.62011-121000-6281201106-0552-05*100190———multicellulartumorspheroidsMCTSs。liquidoverlaymethodHeLaMCTSsMCTSs。。。R73-3Q336A2011-06-302011-07-27No．3097078481171455．1984-．*[e-mail]。1。multicellulartumorspheroidsMCTSs、、MCTSs2～5。。MCTSs。2MCTSs、、。MCTSs。。11.1HeLa-MCTSsHeLa-MCTSs。HeLaDMEM10%1%100mm70%～80%62/125/250/500/1000/9637℃。1.2HeLa-MCTSsHeLa-MCTSsPBS5%。HeLa-MCTSsPBS3%1%。DAPI12hPBS6。1.3HeLa-MCTSsaminoPEGQdot605andcarboxylQdot605invitrogenHeLaHeLa-MCTSs20nMol/L37℃24hPBS。22.1HeLa-MCTSsHeLa-MCTSs2。62/125/250/500/HeLa-MCTSs。62～1000/3～10dHeLa-MCTSs1。1HeLa。Bar=200μmFig．1OpticalimagingofHelacellularspheroids．MCTSsfieldemissionscanningelectronmicroscopyHeLa-MCTSs。MCTSsHeLaHeLa2。HeLa-MCTSsproliferativecellsquiescentcellsapoptoticcells3。2.2HeLa-MCTSs4。2.3HeLa-MCTSsHelaHeLa-MCTSs。HeLa5HeLa-MCTSs5。3HeLa-MCTSs67、。MCTSs。8～10。355J．Chin．Electr．Microsc．Soc．302HeLa-MCTSs250/。Bar=50050202μmFig．2TheexternalmorphologyofintactHelacellularspheroids250cells/well．Bar=50050202μm3HeLa-MCTSs。aHelabcd。Bar=2μmFig．3InternalmorphologyofHeLacellularspheroidsandmonolayercells．aMonolayercellsbProliferativecellscQuiescentcellsdApoptoticcells．Bar=2μmMCTSs。。、11～13。MCTSs。141516。“”。MCTSs。45564HeLa3D5μmBar=75μm。Fig．43-DimagesofHelaspheroids．Thetomo-scanningleftweretakenevery5μmsectionfromthetoptothebottomofanintactspheroidwhilethe3-DimagerightwasreconstructedbyusingthetomographyBar=75μm．5HelaHela。Bar=11μmBar=100μmFig．5QuantumdotspenetrationofHelaHelamonolayercellsBar=11μmandcellularspheroidsBar=100μm．555J．Chin．Electr．Microsc．Soc．301AbbottA．Cellculturebiology'snewdimensionJ．Nature2003424870－872．2FriedrichJSeidelCEbnerRetal．Spheroid-baseddrugscreenconsiderationsandpracticalapproachJ．NatProtoc20094309－324．3FriedrichJEbnerRKunz-SchughartLA．Experimentalanti-tumortherapyin3-Dspheroids-oldhatornewchallengeJIntJRadiatBiol200783849－871．4TimminsNENielsenLK．Generationofmulticellulartumorspheroidsbythehanging-dropmethodJ．MethodsMolMed2007140141－151．5Kunz-SchughartLAKreutzMKnuechelR．Multicellularspheroidsathree-dimensionalinvitroculturesystemtostudytumourbiologyJ．IntJExpPathol1998791－23．6InchWRMcCredieJASutherlandRM．Growthofnodularcarcinomasinrodentscomparedwithmulti-cellspheroidsintissuecultureJ．Growth197034271－282．7SutherlandRM．CellandenvironmentinteractionsintumormicroregionsthemulticellspheroidmodelJ．Science1988240177－184．8HelmchenFDenkW．Deeptissuetwo-photonmicroscopyJ．NatMethods20052932－940．9SidaniMWyckoffJXueCetal．ProbingthemicroenvironmentofmammarytumorsusingmultiphotonmicroscopyJ．JMammaryGlandBiolNeoplasia200611151－163．10VictoraGDSchwickertTAFooksmanDRetal．GerminalcenterdynamicsrevealedbymultiphotonmicroscopywithaphotoactivatablefluorescentreporterJ．Cell2010143592－605．11MichaletXPinaudFFBentolilaLAetal．QuantumdotsforlivecellsinvivoimaginganddiagnosticsJ．Science2005307538－544．12SmithAMNieS．ChemicalanalysisandcellularimagingwithquantumdotsJ．Analyst2004129672－677．13LuccardiniCTribetCVialFetal．SizechargeandinteractionswithgiantlipidvesiclesofquantumdotscoatedwithanamphiphilicmacromoleculeJ．Langmuir2006222304－2310．14KimBHanGToleyBJetal．TuningpayloaddeliveryintumourcylindroidsusinggoldnanoparticlesJ．NatNanotechnol20105465－472．15CarlssonJAckerH．RelationsbetweenpHoxygenpartialpressureandgrowthinculturedcellspheroidsJ．IntJCancer198842715－720．16FolkmanJ．TumorangiogenesisapossiblecontrolpointintumorgrowthJ．AnnInternMed19758296－100．*CorrespondingauthorMulticellularTumorspheroidsMCTSsaversatileinvivo-liketumormodelJIANGQiaoXUEXueMAHui-liZOUGuo-zhangLIANGXing-jie*NationalcenterforNanoscienceandTechnologyBeijing100190ChinaAbstractWepresentaflexibleandhighlyreproduciblemethodforusing3dimensionalmulticellulartumorspheroidsMCTSstoimitatesolidtumorinvivo．WeusedliquidoverlaymethodtocreatetheHelacellularspheroidsandthismodelwasperfectinsimulatingthecellulargradientofproliferativestatus．Theexternalappearanceandinternalcharactersofthe3Dculturedcellswereobservedbyscanningelectronmicroscopyandtransmissionelectronmicroscopy．WeobtainedthetomographyoflivingHelacellularspheroidsandreconstructedthe3Dimagesbymultiphotonmicroscopy．ThepenetrationbehaviorsofquantumdotsandmicellestoHelaspheroidandmonolayercellswereinvestigatedbymultiphotonmicroscopywhichresembledtheprocessofimagingmaterialstransferringintosolidtumorsinvivo．Remarkablyourdataindicatedthe3Dculturesystemandthis3Dimagingmethodcanbeapotential“filter”toscreenoutnanoparticleswithbetterpenetrationfromtheirinherentproperties．AlloftheseapproachesallowedtheexplorationofnanoparticlesandchemotherapeuticsdeliverytosolidtumorinvivotobeeasilyachievedbyapplyingtheMCTSsmodelinvitro．Keywordsmulticellulartumorspheroids3Dculturedcellsquantumdots655