Development of a flow cytometry-based potency assay for measuring the in vitro immunomodulatory properties of mesenchymal stromal cells

Ribeiro, Andreia; Ritter, Thomas; Griffin, Matthew D.; Ceredig, Rhodri

doi:10.1016/j.imlet.2016.07.010

Cited by 16 publications

(20 citation statements)

References 32 publications

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“…It has been shown that MSCs from donors with a high proliferation rate are smaller in size, have longer telomeres and show enhanced ectopic bone forming capacity compared with MSCs from donors with a lower proliferation rate . For other therapeutic applications, different sets of potency tests may be required, such as proposed for the selection of MSC donors with above average immunomodulatory capacity . At the moment, relevant in vitro potency tests that enable selection of MSC donors and batches with enhanced therapeutic efficacy are very limited.…”

Section: Mechanism Of Action; Current State Of the Artmentioning

confidence: 99%

Mesenchymal Stromal Cells Anno 2019: Dawn of the Therapeutic Era? Concise Review

Hoogduijn

Lombardo²

2019

Stem Cells Translational Medicine

125

118

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Summary 2018 was the year of the first marketing authorization of an allogeneic stem cell therapy by the European Medicines Agency. The authorization concerns the use of allogeneic adipose tissue‐derived mesenchymal stromal cells (MSCs) for treatment of complex perianal fistulas in Crohn's disease. This is a breakthrough in the field of MSC therapy. The last few years have, furthermore, seen some breakthroughs in the investigations into the mechanisms of action of MSC therapy. Although the therapeutic effects of MSCs have largely been attributed to their secretion of immunomodulatory and regenerative factors, it has now become clear that some of the effects are mediated through host phagocytic cells that clear administered MSCs and in the process adapt an immunoregulatory and regeneration supporting function. The increased interest in therapeutic use of MSCs and the ongoing elucidation of the mechanisms of action of MSCs are promising indicators that 2019 may be the dawn of the therapeutic era of MSCs and that there will be revived interest in research to more efficient, practical, and sustainable MSC‐based therapies. Stem Cells Translational Medicine 2019;8:1126–1134

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Section: Mechanism Of Action; Current State Of the Artmentioning

confidence: 99%

Mesenchymal Stromal Cells Anno 2019: Dawn of the Therapeutic Era? Concise Review

Hoogduijn

Lombardo²

2019

Stem Cells Translational Medicine

125

118

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“…Flow cytometry is also suitable to analyze cytokine expression in specific cells after intracellular immunostaining, which has already been used to investigate the influence of MSC on T cells [25, 29]. Furthermore, in a possibly trend-setting approach for estimating MSC potency, flow cytometry has been used to analyze MSC-mediated suppression of monocyte activation in whole blood, as determined via intracellular TNF-α staining [31].…”

Section: Introductionmentioning

confidence: 99%

A novel direct co-culture assay analyzed by multicolor flow cytometry reveals context- and cell type-specific immunomodulatory effects of equine mesenchymal stromal cells

Hillmann¹,

Paebst

Brehm³

et al. 2019

PLoS ONE

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The immunomodulatory potential of multipotent mesenchymal stromal cells (MSC) provides a basis for current and future regenerative therapies. In this study, we established an approach that allows to address the effects of pro-inflammatory stimulation and co-culture with MSC on different specific leukocyte subpopulations. Equine peripheral blood leukocyte recovery was optimized to preserve all leukocyte subpopulations and leukocyte activation regimes were evaluated. Allogeneic labeled equine adipose-derived MSC were then subjected to direct co-culture with either non-stimulated, concanavalin A (ConA)-activated or phosphate 12-myristate 13-acetate and ionomycin (PMA/I)-activated leukocytes. Subsequently, production of the cytokines interferon-γ (IFN- γ), interleukin-1 (IL-1) and tumor necrosis factor-α (TNF-α) and presence of FoxP3 were determined in specific cell populations using multicolor flow cytometry. Prostaglandin E2 (PGE2) was measured in the supernatants. ConA-stimulation induced mild activation of leukocytes, whereas PMA/I-stimulation led to strong activation. In T cells, PMA/I promoted production of all cytokines, with no distinct suppressive effects of MSC. However, increased numbers of CD25/FoxP3-positive cells indicated that MSC supported regulatory T cell differentiation in PMA/I-activated leukocyte cultures. MSC also reduced numbers of cytokine-producing B cells and granulocytes, mostly irrespective of preceding leukocyte activation, and reversed the stimulatory effect of ConA on IFN-γ production in monocytes. Illustrating the possible suppressive mechanisms, higher numbers of MSC produced IL-10 when co-cultured with non-stimulated or ConA-activated leukocytes. This was not observed in co-culture with PMA/I-activated leukocytes. However, PGE2 concentration in the supernatant was highest in the co-culture with PMA/I-activated leukocytes, suggesting that PGE2 could still mediate modulatory effects in strongly inflammatory environment. These context- and cell type-specific modulatory effects observed give insight into the interactions between MSC and different types of immune cells and highlight the roles of IL-10 and PGE2 in MSC-mediated immunomodulation. The approach presented could provide a basis for further functional MSC characterization and the development of potency assays.

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“…Assays to assess the immunomodulation capacities of MSC include: Inhibition of mitogen/alloantigen‐induced T‐cell proliferation Suppression of CD154 activation marker expression on CD4 T cells CD200 expression as an indicator or surrogate marker for suppression (however, blocking did not impair suppressive strength) Inducible indoleamine 2,3‐dioxygenase 1 and programmed death ligand 1 expression TNF‐α expression in lipopolysaccharide‐stimulated macrophages MSC‐induced interleukin (IL)‐10 release from blood cells …”

Section: Quality Controlmentioning

confidence: 99%

Production and quality testing of multipotent mesenchymal stromal cell therapeutics for clinical use

Kuçi

Schäfer

2019

Transfusion

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Development of a flow cytometry-based potency assay for measuring the in vitro immunomodulatory properties of mesenchymal stromal cells

Cited by 16 publications

References 32 publications

Mesenchymal Stromal Cells Anno 2019: Dawn of the Therapeutic Era? Concise Review

Mesenchymal Stromal Cells Anno 2019: Dawn of the Therapeutic Era? Concise Review

A novel direct co-culture assay analyzed by multicolor flow cytometry reveals context- and cell type-specific immunomodulatory effects of equine mesenchymal stromal cells

Production and quality testing of multipotent mesenchymal stromal cell therapeutics for clinical use

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