Preparation of mesenchymal stem cells for transplantation: cultivation and quality control

Cellular Therapy and Transplantation

Alexandra A. Aizenstadt¹  Alexandrina S. Khrupina¹  Alexander B. Smolyaninov¹  Irina L. Trofimova¹ 

关键词: mesenchymal stem cells;    cultivation;    quality control;    immunophenotyping;    karyotyping;   

DOI  :  10.3205/ctt-2010-No9-abstract25

学科分类：肿瘤学

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

【 摘 要 】

Mesenchymalstemcells(MSCs)aretheprecursorsofmosthumantissue.Theycanbeefficientlyappliedfortreatmentandpreventionofsuchseriousdiseasesasmyocardialinfarction,diabetes,liverfailure,cardiomyopathy,andseveralautoimmunediseases.HoweverasufficientamountofMCSsarenecessaryforsuccessfulclinicaluse,namely1.5–2.0х106cells/kgofpatientweight.Thisamountisnotpossibletoobtainwithoutcultivation.WeselectedoptimalconditionsforhumanMSCcultivationthatprovidetheirmaximalviabilityandproliferativeactivity.MSCswereculturedonAdvanceStemMedia(Hyclone,NewZealand)for25–40daysfornolongerthan4passages.Thefirstpassagewasonday10–12afterexplantation.Thenthecellculturewasseededevery5–9dayswithaninitialdensityof1.3х103cells/sm2.Inordertocontrolthequalityofcells,cytogeneticanalysisandflowcytometryanalysiswithspecificantibodiestocellsurfacemarkerswereappliedateverypassageononesectionoftheMSCcellculture.Aftereachpassagetheconditionalmediawastestedforsterility.UnambiguousMSCpopulationswereobtained;theirphenotypedoesn’tchangeduringcultivationandisdescribedasCD34-,CD45-,CD44+,CD90+,andCD105+.Inkaryotypingatleast15metaphaseplatespereachsamplewereanalyzed.Noalterationinchromosomestructureoramountwasdetectedinthesamples.Nobacterialneitherfungalcontaminationwasdetectedinthesamples.nbsp;

【 授权许可】

Unknown