Short-Term Withdrawal of Mitogens Prior to Plating Increases Neuronal Differentiation of Human Neural Precursor Cells 英文参考文献.docVIP

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Short-Term Withdrawal of Mitogens Prior to Plating Increases Neuronal Differentiation of Human Neural Precursor Cells 英文参考文献.doc

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Short-Term Withdrawal of Mitogens Prior to Plating Increases Neuronal Differentiation of Human Neural Precursor Cells 英文参考文献

Short-TermWithdrawalofMitogensPriortoPlating IncreasesNeuronalDifferentiationofHumanNeural PrecursorCells TelmaTiemiSchwindt1.,FabianaLouiseMotta1.,GabrielaFilosoBarnabe′1.,CristinaGonc?alves Massant1,AlessanderdeOliveiraGuimara?es2,MariaElisaCalcagnotto1,FabioSilvaConceic?a?o3,Joa?o BoscoPesquero2,StevensRehen3,LuizE.Mello1* 1DepartmentofPhysiology,UniversidadeFederaldeSa?o Paulo(UNIFESP),Sa?o Paulo,Brazil,2DepartmentofBiophysics,UniversidadeFederaldeSa?o Paulo(UNIFESP), Sa?o Paulo,Brazil,3InstitutodeCie?ncias Biome′dicas,UniversidadeFederaldoRiodeJaneiro,RiodeJaneiro,Brazil Abstract Background:Humanneuralprecursorcells(hNPC)arecandidatesforneuraltransplantationinawiderangeofneurological disorders.Recently,muchworkhasbeendonetodeterminehowtheenvironmentforNPCcultureinvitromayaltertheir plasticity.Epidermalgrowthfactor(EGF)andfibroblastgrowthfactor-2(FGF-2)areusedtoexpandNPC;however,itisnot clearifcontinuousexposuretomitogensmayabrogatetheirsubsequentdifferentiation.Hereweevaluatedifshort-term removalofFGF-2andEGFpriortoplatingmayimprovehNPCdifferentiationintoneurons. PrincipalFindings:Wedemonstratethatcultureofneurospheresinsuspensionfor2weekswithoutEGF-FGF-2significantly increasesneuronaldifferentiationandneuriteextensionwhencomparedtocellsculturedusingstandardprotocols.Inthis condition,neuronswerepreferentiallylocatedinthecoreoftheneurospheresinsteadoftheshell.Moreover,afterplating, neuronspresentedradialratherthanrandomlyorientedandlongerprocessesthancontrols,comprisedmostlybyneurons withshortprocesses.Thesechangeswerefollowedbyalterationsintheexpressionofgenesrelatedtocellsurvival. Conclusions: These results show that EGF andFGF-2 removal affects NPC fate and plasticity. Taking into account that a threedimensionalstructureisessentialforNPCdifferentiation,hereweevaluated,forthefirsttime,theeffectsofgrowth factorsremovalinwholeneurospheresratherthaninplatedcellculture. Citation:SchwindtTT,MottaFL,FilosoBarnabe′G,Gonc?alvesMassantC,deOliveiraGuimara?es