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:10012764X(2004)0420246203:R331.1:ACD34+/CD90+X,(/,518020):CD34+3,CD34+,EPICSEliteCD34+/CD90+CD34+83%95%,54%71%,>95%CD34+/CD90+90%,40%50%,>95%CD34+,,CD34+:;;;,CD34+,CD34+,,CD34+/CD90+CD34+/CD38-CD34+/AC133+[1],MiniMACSCD34+,CD34+/CD90+1111:1,2COBE.Spectra.AutoPBSCTM,,:1000ml9g/LNaCl,6002ACD2A,200ml(CD34+),113,45ml/minSysmexSF23000,Beckman2CoulterEPICSEliteCD34+112CD34+MiniMACS(MiltenyiBiotech)CD34+,(10g/LPBS)3;30m,PBS1;300l/108CD34+:100l/108FcRFcCD34+2;100l/5106CD34+CD34+2,,61220min;PBS3,2108/ml30m,LS+/VS+(10ml),,,,CD34+113CD34+/CD90+Beckman2CoulterEPICSElite,,,DelaytimeDriveCD34+CD90,,,CD34+/CD90+,,,26422004224ChineseJournalofClinicalLaboratoryScience,2004,Vol122,No14X:,1969,,,1998,(),©1995-2005TsinghuaTongfangOpticalDiscCo.,Ltd.Allrightsreserved.-www.bbioo.com-www.ebioe.comwww.bbioo.comCD34+83%95%,54%71%,>95%;CD34+/CD90+,CD90+28%40%,,CD34+/CD90+90%,40%50%,>95%(1,13)1123CD34+(%)4.612.343.87CD34+(%)95.4679.2283.71CD34+(%)71.2054.4366.58CD34+/CD90+(%)40.4728.2035.32CD34+/CD90+(%)99.0195.9592.27CD34+/CD90+(%)50.9640.6744.5412CD34+,CD34+,,,,;CD34+/CD90+,,,,,,,453CD34+/CD90+CD34+/CD90-4CD34+5CD34+/CD90+3CD90(Thy21)CD34,,CD34+/CD90+CD34+CD34+/CD90+CD34+,CD34+/CD90+[325]CD34+,(FACS)(CCE),FACS[2]MiltenyiBiotechMACsMiniMACS,50m,,,7422004224ChineseJournalofClinicalLaboratoryScience,2004,Vol122,No14©1995-2005TsinghuaTongfangOpticalDiscCo.,Ltd.Allrightsreserved.-www.bbioo.com-www.ebioe.comwww.bbioo.com,,FACSCD34+/CD90+,,DelaytimeDrive,PMT,,,Drive30000,FACS,2.0107,:[1]BockT,BruggerandW,KanzL(eds).HematopoieticStemCell[M].AlphaMedpress,OnePrestige,Suite290,Miamisburg,Ohio,4534223758USA1997.[2],..:,[M].:,2000120022231[3]ThomelyI,SutherlandDR,NayarR,etal.Replicativestressafterallogeneicbonemarrowtranslation:changescyclingofCD34+CD90+andCD34+CD90-hematopoieticprogenitors[J].Blood,2001,97:187621878.[4]SumikumaT,ShimazakiC,InabaT,etal.CD34+/CD90+cellsinfusedbestpredictlatehaematopoieticreconstitutionfollowingautologousperipheralbloodstemcelltransplantation[J].BrJHaematol,2002,117(1):2382244.[5]PrattG,RawstronAC,EnglishAE,etal.AnalysisofCD34+cellsubsetsinstemcellharvestscanmorereliablypredictrapidityanddurabilityofengraftmentthantotalCD34+celldose,butsteadystatelevelsdonotcorrelatewithbonemarrowreserve[J].BrJHaematol,2001,114(4):9372943.PurificationofCD34+/CD90+progenitorstemcellsusingimmuno2magneticmicrobeadsselectionandflow2cytometricsortinginperipheralbloodleukapheresisXUYong,HUOMei(Dept.ofLaboratoryMedicine,ShenzhenPeoplesHospital,theSecondAffiliatedHospitalofJinanUniversityMedicalCollege,Shenzhen,Guangdong,518020,P.R.ofChina)Abstract:ObjectiveTosetupapracticalmethodforpurificationofCD34+stemcellsanditssubsets.MethodsAliquotsofleukapheresissamplestakenfrom3patientswerefirstlypositive2selectedforCD34+stemcellsbyimmuno2magneticmircobeads,andthenwerepurifiedforCD34+/CD90+progenitorstemcellsusingEPICSElitefluorescence2activatedcellsorter.ResultsThepurityofselectedCD34+stemcellsbyimmuno2magneticseparationwas83%295%withtherecoveryrateof54%271%aswellastheviabilityofmorethan95%.Afterflowcytometricsorting,thepurityandviabilityofCD34+/CD90+progenitorstemcellwere>90%and>95%respectively,witharecoveryof40%250%.ThehighernumbersofCD34+stemcellsinthebeginningapheresishave,thehigherpurityandrecoveryoftheprocedureshave.Themorphologiccharacteristicsbetweenthetwoseparationsweredifferent.ConclusionsWeestablishedaneffectiveprocedureforpurificationofCD34+stemcellsanditssubsetsusingcombinationofimmuno2magneticmircobeadsselectionandfluorescence2activatedflowcytometricsorting,withsatisfiedcellpurity,viabilityandrecovery.Keywords:hematopoieticstemcell;cellseparation;flowcytometry;cellsorting(:2003208202,:2004205206)(:),,,,2004(:20042112012014):,,,(:www.fzzyy.com)11:2004102530(6)21:31:1200(,)41:51:61:0591-2859487,3785575;:0591-3785575;Email:[e-mail]71:20048258422004224ChineseJournalofClinicalLaboratoryScience,2004,Vol122,No14©1995-2005TsinghuaTongfangOpticalDiscCo.,Ltd.Allrightsreserved.-www.bbioo.com-www.ebioe.comwww.bbioo.com