MALDI‐IMS as a Tool to Determine the Myocardial Response to Syndecan‐2‐Selected Mesenchymal Stromal Cell Application in an Experimental Model of Diabetic Cardiomyopathy

Purpose: Mesenchymal stromal cells (MSC) are an attractive tool for treatment of diabetic cardiomyopathy. Syndecan-2/CD362 has been identified as a functional marker for MSC isolation. Imaging mass spectrometry (IMS) allows for the characterization of therapeutic responses in the left ventricle. This study aims to investigate whether IMS can assess the therapeutic effect of CD362+-selected MSC on early onset experimental diabetic cardiomyopathy. Experimental Design: 1 × 106 wild type (WT), CD362−, or CD362+ MSC are intravenously injected into db/db mice. Four weeks later, mice are hemodynamically characterized and subsequently sacrificed for IMS combined with bottom-up mass spectrometry, and isoform and phosphorylation analyses of cardiac titin. Results: Overall alterations of the cardiac proteome signatures, especially titin, are observed in db/db compared to control mice. Interestingly, only CD362+ MSC can overcome the reduced titin intensity distribution and shifts the isoform ratio toward the more compliant N2BA form. In contrast, WT and CD362− MSCs improve all-titin phosphorylation and protein kinase G activity, which is reflected in an improvement in diastolic performance. Conclusions and Clinical Relevance: IMS enables the characterization of differences in titin intensity distribution following MSC application. However, further analysis of titin phosphorylation is needed to allow for the assessment of the therapeutic efficacy of MSC

Cytokine treatment optimises the immunotherapeutic effects of umbilical cord-derived MSC for treatment of inflammatory liver disease

Background: Mesenchymal stromal cells (MSC) possess immunomodulatory properties and low immunogenicity, both crucial properties for their development into an effective cellular immunotherapy. They have shown benefit in clinical trials targeting liver diseases; however the efficacy of MSC therapy will benefit from improvement of the immunomodulatory and immunogenic properties of MSC. Methods: MSC derived from human umbilical cords (ucMSC) were treated for 3 days in vitro with various inflammatory factors, interleukins, vitamins and serum deprivation. Their immunogenicity and immunomodulatory capacity were examined by gene-expression analysis, surface-marker expressions, IDO activity, PGE2 secretion and inhibition of T cell proliferation and IFNγ production. Furthermore, their activation of NK cell cytotoxicity was investigated via CD107a expre

Photoluminescence in Er-doped Ge-As-Se chalcogenide thin films

We report the properties of thermally evaporated Ge11.5As24Se64.5 chalcogenide films ion implanted at energies of 2.25MeV with Erbium at concentrations up to 0.4 mol%. The effect of post implant annealing on the refractive index of the films and on the 4I13/2→ 4I15/2 Er transition was studied. Photoluminescence was found to increase significantly and a lifetime of 1.35 ms was obtained in films annealed at 180oC. Different apparent lifetimes for the 4I13/2→ 4I15/2 transition were obtained for 980nm and 1470nm pumps and the origins of this phenomenon are discussed.Funding information: This research was conducted by the Australian Research Council Centre of Excellence for Ultrahigh bandwidth Devices for Optical Systems (project number CE110001018)

Epigenetic changes in umbilical cord mesenchymal stromal cells upon stimulation and culture expansion.

BACKGROUND Mesenchymal stromal cells (MSCs) are studied for their immunotherapeutic potential. Prior to therapeutic use, MSCs are culture expanded to obtain the required cell numbers and, to improve their efficacy, MSCs may be primed in vitro. Culture expansion and priming induce phenotypical and functional changes in MSCs and thus standardisation and quality control measurements come in need. We investigated the impact of priming and culturing on MSC DNA methylation and examined the use of epigenetic profiling as a quality control tool. METHODS Human umbilical cord-derived MSCs (ucMSCs) were cultured for 3 days with interferon (IFN)γ, transforming growth factor (TGF)β or a multi-factor combination (MC; IFNγ, TGFβ and retinoic acid). In addition, ucMSCs were culture expanded for 14 days. Phenotypical changes and T-cell proliferation inhibition capacity were examined. Genome-wide DNA methylation was measured with Infinium MethylationEPIC Beadchip. RESULTS Upon priming, ucMSCs exhibited a different immunophenotype and ucMSC(IFNγ) and ucMSC(MC) had an increased capacity to inhibit T-cell proliferation. DNA methylation patterns were minimally affected by priming, with only one significantly differentially methylated site (DMS) in IFNγ- and MC-primed ucMSCs associated with autophagy activity. In contrast, 14 days after culture expansion, ucMSCs displayed minor phenotypical and functional changes but showed >4000 significantly DMSs, mostly concerning genes involved in membrane composition, cell adhesion and transmembrane signalling. DISCUSSION These data show that DNA methylation of MSCs is only marginally affected by priming, whereas culture expansion and subsequent increased cellular interactions have a large impact on methylation. On account of this study, we suggest that DNA methylation analysis is a useful quality control tool for culture expanded therapeutic MSCs

Additional file 3: Figure S2. of Cytokine treatment optimises the immunotherapeutic effects of umbilical cord-derived MSC for treatment of inflammatory liver disease

In vivo experimental schemes. (Top) In the first set of experiments mice were treated with CCl4 and 3 hours later pre-treated ucMSC (untreated, IFN-γ, IFN-β, TGFβ, starvation, vitamin B6, Starv + VitB6, RA and MC), which were labeled with Qtracker605 beads, were infused IV. Four hours after ucMSC infusion the mice were sacrificed and prepared for imaging. (Bottom) In the second set of experiments mice were sacrificed 72 hours after CCL4 injection and serum was collected. (TIF 7779 kb

El Diario de Pontevedra : periódico liberal: Ano XXIII Número 6606 - 1906 abril 27

Ex vivo experimental scheme. UcMSC were pre-treated for 3 days. Medium was then refreshed and the cells were left overnight in the incubator. The following day, livers were collected from healthy C57BL/6 mice and directly after isolation cut into slices (diameter = 1 cm and thickness = 150 μm) and placed in the wells on top of the pre-treated ucMSC in the presence of LPS. The liver slices were left overnight in the incubator on a shaker (50 rpm). After 24 hours, supernatant and liver slices were harvested. (TIF 4600 kb