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Cellular Reprogramming Employing Recombinant Sox2 Protein英文文献资料.doc

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Cellular Reprogramming Employing Recombinant Sox2 Protein英文文献资料

HindawiPublishingCorporation StemCellsInternational Volume2012,ArticleID549846,10pages doi:10.1155/2012/549846 ResearchArticle CellularReprogrammingEmployingRecombinantSox2Protein MarcThier,BernhardM¨unst,StephanieMielke,andFrankEdenhofer StemCellEngineeringGroup,InstituteofReconstructiveNeurobiology,UniversityofBonn-Life BrainCenterandHertieFoundation,Sigmund-FreudStra?e25,D-53105Bonn,Germany CorrespondenceshouldbeaddressedtoFrankEdenhofer,f.edenhofer@uni-bonn.de Received1December2011;Accepted23January2012 AcademicEditor:RajarshiPal Copyright?2012MarcThieretal.ThisisanopenaccessarticledistributedundertheCreativeCommonsAttributionLicense, whichpermitsunrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalworkisproperlycited. Inducedpluripotentstem(iPS)cellsrepresentanattractiveoptionforthederivationofpatient-speci?cpluripotentcellsforcell replacementtherapiesaswellasdiseasemodeling.Tobecomeclinicallymeaningful,safeiPScellsneedtobegeneratedexhibitingno permanentgeneticmodi?cationsthatarecausedbyviralintegrationsofthereprogrammingtransgenes.Recently,variousexperi- mentalstrategieshavebeenappliedtoaccomplishtransgene-freederivationofiPScells,includingtheuseofnonintegratingviruses, episomalexpression,orexcisionoftransgenesafterreprogrammingbysite-speci?crecombinasesortransposases.Astraightfor- wardapproachtoinducereprogrammingfactorsisthedirectdeliveryofeithersyntheticmRNAorbiologicallyactiveproteins.We previouslyreportedthegenerationofcell-permeantversionsofOct4(Oct4-TAT)andSox2(Sox2-TAT)proteinsandshowed thatOct4-TATisreprogramming-competent,thatis,itcansubstituteforOct4-encodingvirus.Here,weexploreconditionsfor enhancedSox2-TATproteinstabilizationandfunctionaldeliveryintosomaticcells.Weshowthatcell-permeantSox2proteincan bestabilizedbylipid-richalbuminsupplementsinserumreplacementorlow-serum-supplementedmedia.Employingoptimized conditionsforproteindelivery,wedemonstratethatSox2-TATproteinisabletosubstituteforviralSox2.Sox2-piPScellsexpress pluri