Immunosuppressive capabilities of mesenchymal stromal cells are maintained under hypoxic growth conditions and after gamma irradiation

Nold, Philipp; Hackstein, Holger; Riedlinger, Tabea; Kasper, Cornelia; Neumann, Anne; Mernberger, Marco; Fölsch, Christian; Schmitt, Jan; Fuchs-Winkelmann, Susanne; Barckhausen, Christina; Killer, Madeleine C.; Neubauer, Andreas; Brendel, Cornelia

doi:10.1016/j.jcyt.2014.10.004

Cited by 30 publications

(23 citation statements)

References 53 publications

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“…In order to determine the efficacy of MSC labeling with Au NPs at an optimized concentration of 10 nM, MSCs from eleven different donors were incubated with Au NP for 48 h. MSCs were all in passage 3 to 4, because many cell doublings may impair cell functioning and differentiation [40]. Although the same optimized labeling strategy and Au NP concentration was applied, uptake of Au NPs varied considerably in MSCs derived from different individuals as shown in Fig.…”

Section: Resultsmentioning

confidence: 99%

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Optimizing conditions for labeling of mesenchymal stromal cells (MSCs) with gold nanoparticles: a prerequisite for in vivo tracking of MSCs

Nold¹,

Hartmann²,

Feliu³

et al. 2017

J Nanobiotechnol

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BackgroundMesenchymal stromal cells (MSCs) have an inherent migratory capacity towards tumor tissue in vivo. With the future objective to quantify the tumor homing efficacy of MSCs, as first step in this direction we investigated the use of inorganic nanoparticles (NPs), in particular ca. 4 nm-sized Au NPs, for MSC labeling. Time dependent uptake efficiencies of NPs at different exposure concentrations and times were determined via inductively coupled plasma mass spectrometry (ICP-MS).ResultsThe labeling efficiency of the MSCs was determined in terms of the amount of exocytosed NPs versus the amount of initially endocytosed NPs, demonstrating that at high concentrations the internalized Au NPs were exocytosed over time, leading to continuous exhaustion. While exposure to NPs did not significantly impair cell viability or expression of surface markers, even at high dose levels, MSCs were significantly affected in their proliferation and migration potential. These results demonstrate that proliferation or migration assays are more suitable to evaluate whether labeling of MSCs with certain amounts of NPs exerts distress on cells. However, despite optimized conditions the labeling efficiency varied considerably in MSC lots from different donors, indicating cell specific loading capacities for NPs. Finally, we determined the detection limits of Au NP-labeled MSCs within murine tissue employing ICP-MS and demonstrate the distribution and homing of NP labeled MSCs in vivo.ConclusionAlthough large amounts of NPs improve contrast for imaging, duration and extend of labeling needs to be adjusted carefully to avoid functional deficits in MSCs. We established an optimized labeling strategy for human MSCs with Au NPs that preserves their migratory capacity in vivo.Electronic supplementary materialThe online version of this article (doi:10.1186/s12951-017-0258-5) contains supplementary material, which is available to authorized users.

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Section: Resultsmentioning

confidence: 99%

“…The MCS were cultivated in flasks at 37 °C and 5% CO 2 , until they reached 80% confluence. MSC where used in passages ≤5 due to observed adverse effects on MSC functional capabilities for higher passages as described previously [40]. …”

Section: Methodsmentioning

confidence: 99%

Optimizing conditions for labeling of mesenchymal stromal cells (MSCs) with gold nanoparticles: a prerequisite for in vivo tracking of MSCs

Nold¹,

Hartmann²,

Feliu³

et al. 2017

J Nanobiotechnol

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“…As shown previously, senescence of MSCs is correlated with poor T-cell suppressive capacity [14]. Because senescent MSCs exhibit low metabolic activity [19], we sought to analyze the metabolism of MSCs and PBMCs in coculture in terms of the ECAR (i.e., cellular lactate extrusion as a surrogate of glycolysis) and the OCR (as an indicator of cellular respiration).…”

Section: Resultsmentioning

confidence: 99%

Immunosuppressive capacity of mesenchymal stem cells correlates with metabolic activity and can be enhanced by valproic acid

Killer¹,

Nold²,

Henkenius³

et al. 2017

Stem Cell Res Ther

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BackgroundMesenchymal stem cells (MSCs) have entered the clinic as an Advanced Therapy Medicinal Product and are currently evaluated in a wide range of studies for tissue regeneration or in autoimmune disorders. Various efforts have been made to standardize and optimize expansion and manufacturing processes, but until now reliable potency assays for the final MSC product are lacking. Because recent findings suggest superior therapeutic efficacy of freshly administered MSCs in comparison with frozen cells, we sought to correlate the T-cell suppressive capacity of MSCs with their metabolic activity.MethodsHuman MSCs were obtained from patients’ bone fragments and were employed in coculture with peripheral blood mononuclear cells (PBMCs) in an allogeneic T-cell proliferation assay to measure immunosuppressive function. Metabolic activity of MSCs was measured in real time in terms of aerobic glycolysis quantified by the extracellular acidification rate and mitochondrial respiration quantified by the oxygen consumption rate.ResultsWe show that MSC-induced suppression of T-cell proliferation was highly dependent on individual healthy donors’ lymphocytes. Moreover, coculture with PBMCs increased the glycolytic and respiratory activity of MSCs considerably in a PBMC donor-dependent manner. The twofold to threefold enhancement of cell metabolism was accompanied by higher T-cell suppressive capacities of MSCs. The cryoprotectant dimethyl sulfoxide decreased metabolic and immunosuppressive performances of MSCs while valproic acid (VPA) increased their glycolytic, respiratory and T-cell suppressive capacity.ConclusionsFunctional fitness of MSCs can be determined by measuring metabolic activity and can be enhanced by exposure to VPA. Pretesting the increment of metabolic activity upon interaction of donor MSCs with patient T-cells provides a rational approach for an individualized potency assay prior to MSC therapy.Electronic supplementary materialThe online version of this article (doi:10.1186/s13287-017-0553-y) contains supplementary material, which is available to authorized users.

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“…It has been demonstrated that freezing and thawing affects MSC functionality (Chinnadurai et al, 2016; François et al, 2012; Hoogduijn et al, 2016; Kadekar et al, 2016; Moll et al, 2014, 2016; Nold et al, 2015; Pogozhykh et al, 2017; Pollock et al, 2015; Quimby et al, 2013). Here we further show that thawed MSCs display a defective secretome signature with T cell proliferation.…”

Section: Discussionmentioning

confidence: 99%

Potency Analysis of Mesenchymal Stromal Cells Using a Combinatorial Assay Matrix Approach

et al. 2018

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SUMMARY Assays that can characterize MSC immune potency need to be identified for use in advanced clinical trials. MSCs possess a number of putative regenerative and immunomodulatory properties, and an assay matrix approach may best capture involved effector pathways. We have tested two assay systems to measure the potency of MSCs derived from human subjects: MSC secretome analysis and a quantitative RNA-based array for genes specific to immunomodulatory and homing properties of MSCs. Secretome analysis identified a unique cytokine signature that is upregulated by MSCs or downregulated in responder PBMCs and correlated with T cell suppression. Use of interferon-γ as a surrogate for the action of activated PBMCs on MSCs served as an alternative for the use of human PBMCs as responder cells in a potency assay. Our approach and results define and simplify the multifunctional or matrix responses of MSCs and may serve as a platform for robust potency analysis.

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Immunosuppressive capabilities of mesenchymal stromal cells are maintained under hypoxic growth conditions and after gamma irradiation

Cited by 30 publications

References 53 publications

Optimizing conditions for labeling of mesenchymal stromal cells (MSCs) with gold nanoparticles: a prerequisite for in vivo tracking of MSCs

Optimizing conditions for labeling of mesenchymal stromal cells (MSCs) with gold nanoparticles: a prerequisite for in vivo tracking of MSCs

Immunosuppressive capacity of mesenchymal stem cells correlates with metabolic activity and can be enhanced by valproic acid

Potency Analysis of Mesenchymal Stromal Cells Using a Combinatorial Assay Matrix Approach

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