简介：backgroundBonemarrowmesenchymalstemcells(BMSCs)canbeisolatedandculturedtomanypassages.However,StemcellsincludingBMSCsquicklyundergosenescenceinculture.Thecellsenescenceandmulti-directionaldifferentiationhavehamperedproducingBMSCsinquantitywiththeirundifferentiatedstate.Inthisstudywereportanaturalcompound,vitaminC(Vc),maintainsBMSCsstemproperty.MethodsHumanBMSCswereisolatedfrombonemarrowandpurifiedby1.073g/mLdensitygradientcentrifugation.50ng/mLVcwereaddedtoBMSCsfordifferenttimepoint.FlowcytometrywasusedtodetectcellsurfacemarkersofBMSCswithorwithoutVctreatment.BMSCsproliferationwasanalyzedbyMTTassay.PCR(polymerasechainreaction)andreal-timePCRwereusedfordetectingc-kit,nanog,andOct-4genesexpressionlevels.DNAmethyltransferase(Dnmt)1andDnmt3blevelswerealsodetectedbyreal-timePCR.ResultsFlowcytometryshowedthatafterVctreatmentfor6h,thesurfacemarkersofBMSCswerealmostunchanged.VcincreasedtheproliferationactivityofBMSCsfrom6hto24h.PCRshowedtheexpressionofc-kit,nanog,andoct-4geneswereobviouslyincreasedinVctreatedgroupthancontrolgroupat12h.Real-timePCRshowedthatthelevelofc-kit,nanog,andoct-4geneswereunregulatedfrom6hto12hcomparedwithcontrolgroup.VcalsoincreasedDnmt3bbutnotDnmt1geneexpression.ConclusionsOurresultsshowedVcactsatleastacceleratesBMSCsproliferationandmaintainsstemcellproperty.Inourstudy,wehighlightedamethodofimprovingthespeedofBMSCsgenerationandprovidedadditionalinsightsintothemechanisticbasisofpreventingBMSCssenescence.

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简介：BackgroundCurrentlyusedheartvalveprosthesesareassociatedwithanticoagulationcomplicationsorlimiteddurability.Theadvancementofstemcellstudyandtissue-engineeredheartvalveresearchmayofferarelativelyidealsolutiontotheseproblems.MethodsBonemarrowwasaspiratedfromsternumoflambgoatstoisolateBMCs.Cellswereidentifiedbyflowcytometryanditscapacityofdifferentiation.CellularviabilitywasassessedwithRhdomine123staining.1×107cellswereseededonapatchofPGAsheet.Aftertwo-dayinvitroculture,theautologouscell/scaffoldsheetswereusedtoreplacetherightposteriorpulmonaryvalveleafletsundercardiopulmonarybypass.Theleafletswereexplantedat2days,2,6,8and10weeksafterimplantation.Thesampleswereexaminedmacroscopically,histologically,immunohistochemically,andbyScanningElectronMicroscope(SEM).Twogoatswereimplantedwithacellularsheetsandestablishedasacontrolgroup.ResultsBMCsexhibitedfibroblastoidmorphologywithgoodviability.FlowcytometryshowednegativeCD14andCD45expression.InvitroculturedBMCsdemonstratedthepotentialtodifferentiateintoadipocytes.Theexplantedleafletsresembledthecharacteristicsofnativeleafletsmacroscopicallyinthecellulargroup.Histologyshowedextracellularmatrixwassynthesizedandcellsweredistributedinthesingle-layeredleaflets.ImmunohistochemistryrevealedpositivestainingforvonWillebrandfactor,α-SMA,vimentin.AconfluentcellsurfacewasformedontheexplantedTEHLs.Nocalciumdepositedontheleaflets.Incontrolgroup,theacellularscaffoldswerecompletelydegraded,withoutleafletremainedat8weeks.ConclusionsItispossibletocreatetissue-engineeredheartvalvesinvivousingautologousbonemarrow-derivedcells.

简介：Intherecentpast,bonemarrow(BM)-derivedcellshavebeenusedtoregeneratedamagedcardiovasculartissuespostmyocardialinfarction(MI).Recentclinicaltrialshaveshowncontroversialresultsinrecoveringdamagedcardiactissue.Newprogresshasshownthattheunderlyingmechanismsofcell-basedtherapyreliesmoreheavilyonhumoralandparacrineeffectsratherthanonnewtissuegeneration.However,studieshavealsoreportedthepotentialofnewendothelialcellgenerationfromBMcells.Thus,effortshavebeenmadetoidentifycellshavinghigherhumoralortherapeuticeffectsaswellastheirsurfacemarkers.Specifically,BM-derivedCD31~+cellswereisolatedbyasurfacemarkeranddemonstratedhighangio-vasculogeniceffects.IwillpresentrecentadvancesinthetherapeuticuseofBM-derivedcellsandtheusefulnessofCD31~+cellsasanextgenerationcelltherapy.

简介：BackgroundOurpreviousstudyshowedthe150mg/mLfetalcardiacsupernatant(FCS)couldinducedifferentiationofBMSCsintocardiomyocye-likecellswithoutcardiomyocytetouch,butdifferentiationefficiencyisnothighenough.Inhibitionofglycogensynthasekinase-3enhancedtheproliferationandsurvivesofstemcells.Wetestedif6-bromoindirubin-3-oxime(BIO,glycogensynthasekinase-3inhibitor)enhancestheeffectsofFCSondifferentiationofBMSCsandexplorethegrowthfactorsinFCS.MethodsBMSCswereisolatedfromthefemurandtibiaoffour-week-oldmaleSprague-Dawleyratsandco-culturedwithFCS(150mg/mL)thatwasmadefromfetalheartsfromnineteen-daypregnantWistarrats.BIOwithdifferentconcentration(0,1,10,and100nM)wasintroducedinculturedishes.Transforminggrowthfactorbeta1(TGF-β1),bonemorphogeneticprotein2(BMP-2)andAktincardiacsupernatantandculturemediumwereassayedwithELISAmethods.ResultsAfterco-culturingwithFCS,beatingmyotubeswereobservedin25.9%BMSCsdishesafter1to2weeks’culture.ThelevelsofTGF-β1andBMP-2inFCSconcentrationswerenomorethanthatinyoungandadultcardiacsupernatant.AllBIOgroupssignificantlyenhancedtheeffectsofFCSondifferentiationofBMSCsintothecardiomyocyte-likecells(1nM,83%;10nM,73%;100nM,100%).AktlevelswerehigherinBMSCsculturalmediumwithFCS.ConclusionsFCScouldinducethedifferentiationofBMSCsintothecardiomyocyte-likecells.TGF-β1andBMP-2mightnotplayaroleinthedifferentiationofBMSCsinducedbyFCS.BIOenhancedtheeffectsofFCSonthedifferentiationofBMSCsintocardiomyocyte-likecells,whichmightinvolvetheAktpathway.