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Cytokine secretion profiling of human mesenchymal stem cells by antibody array.

ABSTRACT: Mesenchymal stem cells (MSCs) provide not only cell sources for connective tissues but also the control of hematopoiesis and immune response. A multitude of cytokines and growth factors secreted from MSCs are known to confer such multifunctional functionality, but their overall spectrum and the respective expression strength have not been thoroughly illustrated. In this study, we have obtained the comprehensive cytokine secretion profile of human bone marrow (BM)-derived MSCs, with the use of an antibody array recognizing 120 cytokines and chemokines. The array membrane incubated with the secretion media of the cells featured a predominant hybridization signal for IL-6 and moderately elevated signals for IL-8, TIMP-2, MCP-1, VEGF and OPG. This cytokine secretion profile was found to be common to all cell lines from three different donors, and also similar but not identical to that of umbilical cord blood-derived cells, suggesting that the trophic nature of the MSCs might depend slightly on the cell origin but not on individuality of the donors. Our results here may provide the molecular basis for further studies on MSC-assisted biological processes, such as connective tissue homeostasis, hematopoiesis and immune modulation.

SUBMITTER: Park CW

PROVIDER: S-EPMC4021795 | biostudies-literature | 2009 May

REPOSITORIES: biostudies-literature

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Cytokine secretion profiling of human mesenchymal stem cells by antibody array.

Park Chae Woon CW Kim Keun-Soo KS Bae Sohyun S Son Hye Kyeong HK Myung Pyung-Keun PK Hong Hyo Jeong HJ Kim Hoeon H

International journal of stem cells 20090501 1

Mesenchymal stem cells (MSCs) provide not only cell sources for connective tissues but also the control of hematopoiesis and immune response. A multitude of cytokines and growth factors secreted from MSCs are known to confer such multifunctional functionality, but their overall spectrum and the respective expression strength have not been thoroughly illustrated. In this study, we have obtained the comprehensive cytokine secretion profile of human bone marrow (BM)-derived MSCs, with the use of an ...[more]

PMID: 24855521

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Project description:BackgroundThe paracrine effects of multipotent mesenchymal stromal cells (MSCs) are mediated by their secretome composed by soluble factors (i.e., cytokines, growth factors, hormones) and extracellular vesicles (EVs). EVs promote intercellular communication, and the EV cargoes [e.g., proteins, soluble factors, microRNAs (miRNAs), messenger RNA (mRNA), DNA] reflect the molecular and functional characteristics of their parental cells. MSC-derived EVs (MSC-EVs) are currently evaluated as subcellular therapeutics. A key function of the MSC secretome is its ability to promote immune tolerance (i.e., immunopotency), a property that is enhanced by priming approaches (e.g., cytokines, hypoxia, chemicals) and inversely correlates with the age of the MSC donors. We evaluated mechanisms underlying MSC vesiculation and the effects of inflammation and aging on this process.MethodsWe evaluated the effects of interferon gamma (IFN-γ) and tumor necrosis factor alpha (TNF-α) on human adipose-derived MSC: (a) vesiculation (custom RT2 Profiler PCR Array), (b) EV profiles (Nanoparticle Tracking Analysis and Nanoparticle Flow Cytometry), (c) EV cargo (proteomic analysis and Western blot analysis), and (d) immunopotency (standard MSC:CD4 T cell proliferation inhibition assay). We confirmed the role of RAB27B on MSC vesiculation (RAB27B siRNA) and assessed its differential contribution to vesiculation in adult and pediatric MSCs (qPCR).ResultsCytokine priming upregulated RAB27B in adipose-derived MSCs increasing their secretion of exosome-like small EVs (sEVs; < 200 nm) containing two key mediators of immunopotency: A20 and TSG-6. These EVs inhibited T cell proliferation in a dose-dependent manner. RAB27B siRNA inhibited MSC vesiculation. Adipose-derived MSCs isolated from pediatric donors exhibited higher RAB27B expression and secreted more sEVs than adult MSCs.ConclusionsCytokine priming is a useful strategy to harvest anti-inflammatory MSC-sEVs for clinical applications. Of relevance, donor age should be considered in the selection of MSC-sEVs for clinical applications.

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Cytokine secretion profiling of human mesenchymal stem cells by antibody array.

Publications

Cytokine secretion profiling of human mesenchymal stem cells by antibody array.

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