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Improved knowledge of the immunological properties of mesenchymal stem cells(MSCs) creates a potential cell-mediated immunotherapeutic approach for arthriticdiseases. The low frequency of MSCs necessitates their in vitro expansion priorto clinical use. As sequential passage has been used as the most popular strategyfor expansion of MSCs, the effect of long-term culture on the immunological propertiesof MSCs is not clear. In this study, we observed that the morphology of MSCs showedthe typical characteristics of the Hayflick model of cellular aging during sequentialexpansion. The growth kinetics of MSCs decreased while the number of MSCs stainingpositive for SA β-gal (senescence marker) increased in long-term culture. Althoughlong-term culture exerts less of an effect on the immunophenotype of MSCs, theimmunosuppressive effects of MSCs on the allogeneic T-cell proliferation, activation-antigenexpression (CD69 and CD25) and cytokine production (IFN-γ, TNF-α, IL-10) weresignificantly impaired following stimulation with phytohemagglutinin (PHA).

Long-term culture in vitro impairs the immunosuppressive activity of mesenchymal stem cells on T cells