Erythroid Progenitor Cells Expanded from Peripheral Blood without Mobilization or Preselection Molecular Characteristics and Functional Competence 英文参考文献.docVIP

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Erythroid Progenitor Cells Expanded from Peripheral Blood without Mobilization or Preselection Molecular Characteristics and Functional Competence 英文参考文献.doc

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Erythroid Progenitor Cells Expanded from Peripheral Blood without Mobilization or Preselection Molecular Characteristics and Functional Competence 英文参考文献

ErythroidProgenitorCellsExpandedfromPeripheral BloodwithoutMobilizationorPreselection:Molecular CharacteristicsandFunctionalCompetence ClaudiaFilippone1*¤,RauliFranssila1,ArunKumar1,LeenaSaikko2,PanuE.Kovanen2,MariaSo¨derlund- Venermo1,KlausHedman1 1DepartmentofVirology,HaartmanInstitute,UniversityofHelsinkiandHelsinkiUniversityLaboratoryDivision,Helsinki,Finland,2DepartmentofPathology,Haartman Institute,UniversityofHelsinki,Helsinki,Finland Abstract Background:Continueddevelopmentofin-vitroproceduresforexpansionanddifferentiationoferythroidprogenitorcells (EPC)isessentialnotonlyinhematologyandstemcellresearchbutalsovirology,inlightofthestricterythrotropismofthe clinicallyimportanthumanparvovirusB19. Methodology/Principal Findings: We cultured EPC directly from ordinary blood samples, without ex vivo stem cell mobilizationorCD34+cellinvitropreselection.Profoundincreaseintheabsolutecellnumberandclusteringactivitywere observedduringculture.ThecellsobtainedexpressedtheEPCmarkercombinationCD36,CD71andglycophorin,butnone ofthelymphocyte,monocyteorNKmarkers.ThefunctionalityofthegeneratedEPCwasexaminedbyaninvitroinfection assaywithhumanparvovirusB19,tropicforBFU-EandCFU-Ecells.Followinginfection(i)viralDNAreplicationandmRNA productionwereconfirmedbyquantitativePCR,and(ii)structuralandnonstructuralproteinswereexpressedin.50%of thecells.Astheoverallcellnumberincreased100–200fold,andtheproportionofcompetentEPC(CD34+toCD36+ )rose from ,0.5% to .50%, the in vitro culture procedure generated the EPC at an efficiency of .10 000-fold. Comparative culturingofunselectedPBMCandexvivo-preselectedCD34+cellsproducedqualitativelyandquantitativelysimilaryieldsof EPC. Conclusions/Significance:ThisapproachyieldingEPCdirectlyfromunmanipulatedperipheralbloodisgratifyinglyrobust andwillfacilitatethestudyofmyeloidinfectiousagentssuchastheB19virus,aswellastheexaminationoferythropoiesis anditscellularandmolecularmechanisms. Citation:FilipponeC,FranssilaR,KumarA,SaikkoL,KovanenPE,etal.(2010)Eryt