Minimal-criteria-for-defining-multipotent-mesenchy

POSITIONPAPERMinimalcriteriafordefiningmultipotentmesenchymalstromalcells.TheInternationalSocietyforCellularTherapypositionstatementMDominici1,KLeBlanc2,IMueller3,ISlaper-Cortenbach4,FCMarini5,DSKrause6,RJDeans7,AKeating8,DJProckop9andEMHorwitz101LaboratoryofCellBiologyandAdvancedCancerTherapy,Oncology-HematologyDepartment,UniversityofModenaandReggioEmilia,Modena,Italy,2CenterforAllogeneicStemCellTransplantation,DepartmentofLaboratoryMedicine,KarolinskaUniversityHospital,KarolinskaInstitute,Stockholm,Sweden,3UniversityChildren’sHospital,DepartmentofHematologyandOncology,Tuebingen,Germany,4DepartmentofMedicalImmunology,UMCUtrecht,Utrecht,theNetherlands,5DepartmentofBloodandMarrowTransplant,UT-MDAndersonCancerCenter,Houston,Texas,USA,6DepartmentofLaboratoryMedicine,YaleUniversitySchoolofMedicine,NewHaven,Connecticut,USA,7AthersysInc.,Cleveland,Ohio,USA,8DepartmentofMedicalOncologyandHematologyPrincessMargaretHospital/OntarioCancerInstituteToronto,Ontario,Canada,9CenterforGeneTherapy,TulaneUniversityHealthSciencesCenter,NewOrleans,Louisiana,USAand10DivisionsofStemCellTransplantationandExperimentalHematology,StJudeChildren’sResearchHospital,MemphisTennessee,USATheconsiderabletherapeuticpotentialofhumanmultipotentmesench-ymalstromalcells(MSC)hasgeneratedmarkedlyincreasinginterestinawidevarietyofbiomedicaldisciplines.However,investigatorsreportstudiesofMSCusingdifferentmethodsofisolationandexpansion,anddifferentapproachestocharacterizingthecells.Thusitisincreasinglydifficulttocompareandcontraststudyoutcomes,whichhindersprogressinthefield.Tobegintoaddressthisissue,theMesenchymalandTissueStemCellCommitteeoftheInternationalSocietyforCellularTherapyproposesminimalcriteriatodefinehumanMSC.First,MSCmustbeplastic-adherentwhenmaintainedinstandardcultureconditions.Second,MSCmustexpressCD105,CD73andCD90,andlackexpressionofCD45,CD34,CD14orCD11b,CD79aorCD19andHLA-DRsurfacemolecules.Third,MSCmustdifferentiatetoosteoblasts,adipocytesandchondroblastsinvitro.Whilethesecriteriawillprobablyrequiremodificationasnewknowledgeunfolds,webelievethisminimalsetofstandardcriteriawillfosteramoreuniformcharacterizationofMSCandfacilitatetheexchangeofdataamonginvestigators.KeywordsMSC,stemcells,adherentcells,immunophenotype,differentiation.BiologicandclinicalinterestinMSChasrisendramati-callyoverthelasttwodecades,asshownbytheever-increasingnumberofresearchteamsstudyingthesecells.NotonlyareestablishedlaboratoriesfocusingonMSCbutnewinvestigatorsarerapidlybeingattractedtothefield,whichwillundoubtedlyacceleratescientificdiscoveryandthedevelopmentofnovelcellulartherapies.However,thissoaringinteresthasalsogeneratedmanyambiguitiesandinconsistenciesinthefield.Tobegintoaddresstheseissues,arecentreportfromtheInternationalSocietyforCellularTherapy(ISCT)statedthat‘multipotentmesenchymalstromalcells’(MSC)isthecurrentlyrecommendeddesignation[1]fortheplastic-adherentcellsisolatedfromBMandothertissuesthathaveoftenbeenlabeledmesenchymalstemcells[2].ThedefiningcharacteristicsofMSCareinconsistentamonginvestigators.ManylaboratorieshavedevelopedmethodstoisolateandexpandMSC,whichinvariablyhavesubtle,andoccasionallyquitesignificant,differences.Furthermore,investigatorshaveisolatedMSCfromavarietyoftissueswithostensiblysimilarproperties[3].ThesevariedtissuesourcesandmethodologiesofcellCorrespondenceto:MassimoDominici,MD,LaboratoryofCellBiologyandAdvancedCancerTherapy,Oncology-HematologyDepartment,UniversityofModenaandReggioEmilia,ViaDelPozzo,71Modena41100,[e-mail](2006)Vol.8,No.4,315
317–2006ISCTDOI:10.1080/14653240600855905preparationbegthequestionofwhethertheresultingcellsaresufficientlysimilartoallowforadirectcomparisonofreportedbiologicpropertiesandexperimentaloutcomes,especiallyinthecontextofcelltherapy.Thisquestionofcellequivalenceis,inpart,becauseofthelackofuniversallyacceptedcriteriatodefineMSC.Mostim-portantly,theinabilitytocompareandcontraststudiesfromdifferentgroupsislikelytohinderprogressinthefield.Toaddressthisproblem,theMesenchymalandTissueStemCellCommitteeoftheISCTproposesasetofstandardstodefinehumanMSCforbothlaboratory-basedscientificinvestigationsandforpre-clinicalstudies.Theseidentifyingcriteriashouldnotbeconfusedwithreleasespecificationsforclinicalstudies,asthecurrentproposalisintendedsolelyasidentifyingcriteriaforresearchpur-poses.Theaimofthispositionstatementistoprovidethescientificcommunitywithastandardsetofcriteria,basedonthebestcurrentlyavailabledata,todefinetheidentityofMSC,recognizingthatfutureresearchwillprobablymandatearevisionofthecriteriaasnewdataemerge.WeproposethreecriteriatodefineMSC:/adherencetoplastic/specificsurfaceantigen(Ag)expression/multipotentdifferentiationpotential(Table1).First,MSCmustbeplastic-adherentwhenmaintainedinstandardcultureconditionsusingtissuecultureflasks.Second,]/95%oftheMSCpopulationmustexpressCD105,CD73andCD90,asmeasuredbyflowcytometry.Additionally,thesecellsmustlackexpression(5/2%positive)ofCD45,CD34,CD14orCD11b,CD79aorCD19andHLAclassII.Third,thecellsmustbeabletodif