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

Abstract

Abstract. Improved knowledge of the immunological properties of mesenchymal stem cells (MSCs) creates a potential cell-mediated immunotherapeutic approach for arthritic diseases. The low frequency of MSCs necessitates their in vitro expansion prior to clinical use. As sequential passage has been used as the most popular strategy for expansion of MSCs, the effect of long-term culture on the immunological properties of MSCs is not clear. In this study, we observed that the morphology of MSCs showed the typical characteristics of the Hayflick model of cellular aging during sequential expansion. The growth kinetics of MSCs decreased while the number of MSCs staining positive for SA β-gal (senescence marker) increased in long-term culture. Although long-term culture exerts less of an effect on the immunophenotype of MSCs, the immunosuppressive effects of MSCs on the allogeneic T-cell proliferation, activation-antigen expression (CD69 and CD25) and cytokine production (IFN-γ, TNF-α, IL-10) were significantly impaired following stimulation with phytohemagglutinin (PHA).

SDF-1 overexpression by mesenchymal stem cells enhances GAP-43-positive axonal growth following spinal cord injury

“…In this study, high-passage MSCs were used, as they have been shown to produce greater treatment effects than low-passaged cells in rodent models of neurodegenerative diseases (Rossignol et al, 2015). This relationship, however, may not be consistent across different neurological pathologies, and/or species (Li et al, 2012).…”

Assessment of the Immunomodulatory Properties of Human Mesenchymal Stem Cells (MSCs)

“…Increasing reports demonstrate that in vitro expansion of MSCs result in senescence of these primary stem cells, and that the differentiation capacity of these stem cells is altered or compromised 22,24 . While currently there are only a few published report with regards to the effects of senescence on MSC immunomodulation 22 , caution should be taken when using MSCs which have been passaged for a prolonged period of time based on our own experience as well (data not shown). Senescence is manifested as a lack of proliferation without cell death, and flattened, enlarged morphology (youthful MSCs are fibroblastic in shape).…”

“…One other important aspect which can affect the reproducibility of these assays is the passage number of the MSCs utilized. It has been noted that the mere act of removing MSCs from their physiologic in vivo environment and cryopreservation can compromise function, including immunomodulation 22,23 . Increasing reports demonstrate that in vitro expansion of MSCs result in senescence of these primary stem cells, and that the differentiation capacity of these stem cells is altered or compromised 22,24 .…”

“…It has been noted that the mere act of removing MSCs from their physiologic in vivo environment and cryopreservation can compromise function, including immunomodulation 22,23 . Increasing reports demonstrate that in vitro expansion of MSCs result in senescence of these primary stem cells, and that the differentiation capacity of these stem cells is altered or compromised 22,24 . While currently there are only a few published report with regards to the effects of senescence on MSC immunomodulation 22 , caution should be taken when using MSCs which have been passaged for a prolonged period of time based on our own experience as well (data not shown).…”

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“…Mesenchymal stem cells lack expression of major histocompatibility complex class II and costimulatory molecules such as CD80, CD86, or CD40. [29][30][31] In addition, human amniotic mesenchymal stromal cells may modulate immune cell activities and may be transplanted across major histocompatibility complex barriers. [32][33] Amniotic cells may suppress T-cell proliferation because of cell-cell contact and secrete of cytokines.…”