Biological properties of human bone marrow mesenchymal stem cells (MSCs) during ex vivo expansion for clinical use

Cellular Therapy and Transplantation

Tatiana V. Shamanskaya¹  Bazarma B. Purbueva¹  Sergey A. Roumiantsev¹  Elena Yu. Osipova¹  Andrey Yu. Yustugov¹ 

关键词: mesenchymal stem cells;    bone marrow;    ex vivo expansion;    immunophenotype;    genetic stability;   

DOI  :  10.3205/ctt-2010-No9-abstract66

学科分类：肿瘤学

来源: Universitaetsklinikum Hamburg - Eppendorf / University Medical Center Hamburg - Eppendorf

【 摘 要 】

Objectives:MSCsaremultipotentcellswithmanypotentialclinicalapplications.However,thelackofaprecisedefinitionofcellpreparationandtheheterogeneityoftheobtainedproductcallforvalidationcriteriaofthefinalproductoftheexpansion.Aims:Toestimateimmunophenotypeandgeneticstability(karyotypeandaneuploidcellfrequency)ofMSCsfromhealthydonorsduringexvivoexpansion.Methods:TheMSCs(n=70)werepreparedbyseedingbonemarrowMNCinflasksinlowglucoseDMEMwith20%FCS.Theimmunophenotypeoftheculturedcellswasanalyzed:CD90,CD105,CD73,CD45,CD31,CD34,CD133.ThekaryotypewasanalyzedbyG-bandingtechnique.Toanalyzethelevelofaneuploidy,fluorescentinsituhybridization(FISH)withchromosomeenumerationprobe(CEP)studieswereperformed.Results:InallcasesthekaryotypeoftheMSCsduringexpansionremainedstable:46,XYor46,XX.Thedisomyratefor6and8chromosomesintheMSCcultureswas98%,andthetetrasomyratewas0.9%anddidnotchangeduringcultivation.Themonosomyrateforsexchromosomes(XandY)intheMSCcultureswashigherthantherateoftrisomy(p≤0.05).TherateofaneuploidyintheMSCculturesdidnotchangeduringcultivation.FISH-analysisofMSCrevealed3sampleswithabnormalclones(1withtrisomy8,and2withmonosomyX).Theseclonesaroseonearlypassagesandremainedintheculturesduringexpansion.Conclusions:Duringtheculturingprocedureschromosomeabnormalitiescanarisewhichcanleadtolongtermconsequencesofcelltherapy.FurtherresearchisrequiredconcerningdifferentpassagingtechniquesandcultureconditionstobetterunderstandtheireffectsonMSCchromosomalstability.

【 授权许可】

Unknown