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1、GeneTherapy(2009)16,154–158&2009MacmillanPublishersLimitedAllrightsreserved0969-7128/09$32.00www.nature.com/gtSHORTCOMMUNICATIONAnefficienttransfectionmethodformouseembryonicstemcellsBSKo1,2,5,TCChang1,5,SKShyue3,YCChen4andJYLiou11NationalHealthResearchInstitute
2、s,ZhunanTown,MiaoliCounty,Taiwan;2DepartmentofInternalMedicine,NationalTaiwanUniversityHospital,Taipei,Taiwan;3InstitutesofBiomedicalScience,AcademicSinica,Taipei,Taiwanand4DepartmentofLaboratoryMedicine,NationalTaiwanUniversityHospital,Taipei,TaiwanEmbryonicst
3、em(ES)cellsareconsideredtohavepotentialsOurresultsshowthattransfectionbyEffecteneachievesthefortissueregenerationandtreatmentofdiversehumanefficiencyof498%inCCEand480%inD3cells.Thediseases.EScellsarecapableofindefiniterenewalandoptimalratioofDNA:EffecteneforEGFPt
4、ransfectionisproliferation,whichcanbeinducedtodifferentiateintotissuesbetween1:4and1:8.Transient-expressedEGFPorendo-ofallthreegermlines.Despitetheseexcitingpotential,itgenousproteinkinaseA(PKA)weresignificantlyknockedremainsunclearastohowtherenewalanddifferenti
5、ationdownbyEffectenetransfectionofspecificsiRNA.HighEGFPprogramsareoperatedandregulatedatthegeneticlevel.levelexpressionandaccumulationinmEScellsinducesGeneticmanipulationsuchasdeliveryofexogenousgeneminorcytotoxicitybutcanbereducedbyintroducingsiRNAofexpression
6、orknockdownwithsmallinterferingRNA(siRNA)EGFP.Further,thistransfectionmethoddidnotsignificantlyiscommonlyusedinmostofcancerortransformedcellsbutaffectmESpropertiesofproliferationordifferentiation.OurrelativelyrareinEScells.Inthisstudy,wecomparetheresultsprovidea
7、noptimalprotocoltoachieveanefficienttransfectionefficaciesofseveralliposome-basedtransfectiontransfectionformEScells.methodsbyintroductionofaplasmidencodingenhancedGeneTherapy(2009)16,154–158;doi:10.1038/gt.2008.125;greenfluorescentprotein(EGFP)intomouseES(mES)cel
8、ls.publishedonline31July2008Keywords:mouseembryonicstemcells;transfection;efficiency;greenfluorescentprotein;smallinterferingRNAMouseembryonicstem(mES)cellsderivedfromdescribe
