Vitamin C maintaining bone marrow mesenchymal stem cell self-renewal

(整期优先)网络出版时间:2011-01-11
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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.