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dc.contributor.authorRibeiro, Andreia
dc.contributor.authorRitter, Thomas
dc.contributor.authorGriffin, Matthew
dc.contributor.authorCeredig, Rhodri
dc.date.accessioned2017-02-13T12:06:50Z
dc.date.issued2016-07-20
dc.identifier.citationRibeiro, Andreia, Ritter, Thomas, Griffin, Matthew, & Ceredig, Rhodri. (2016). Development of a flow cytometry-based potency assay for measuring the in vitro immunomodulatory properties of mesenchymal stromal cells. Immunology Letters, 177, 38-46. doi: http://dx.doi.org/10.1016/j.imlet.2016.07.010en_IE
dc.identifier.issn1879-0542
dc.identifier.urihttp://hdl.handle.net/10379/6288
dc.description.abstractHuman bone marrow-derived mesenchymal stromal/stem cells (MSC) have well-documented modulatory effects on multiple immune cell types. Although these effects are linked to their therapeutic benefit in diverse diseases, a reliable, quantitative assay of the immunomodulatory potency of individual human MSC preparations is lacking. The aims of this study were to develop an optimised rapid turnaround, flow cytometry-based whole-blood assay to monitor MSC potency and to validate its application to MSC immunomodulation. A protocol for short-term LPS stimulation of anti-coagulated whole blood samples followed by combined surface CD45/CD14 and intracellular TNF-alpha staining was initially developed for analysis on a 4 colour desktop cytometer. Optimal monocyte activation was dependent on the presence of extracellular calcium ions thereby precluding the use of EDTA and sodium citrate as anticoagulants. Optimal assay conditions proved to be 1 ng/mL ultrapure-LPS added to 10-fold diluted, heparin anti coagulated whole blood incubated for 6h at 37 degrees C. Under these conditions, addition of human bone marrow-derived MSC (hBM-MSC) from multiple donors resulted in a reproducible, dose-dependent inhibition of LPS-stimulated monocyte TNF-a expression. We conclude that this protocol represents a practical, quantitative assay of a clinically relevant functional effect of hBM-MSCs as well as other immunomodulatory agents. (C) 2016 European Federation of Immunological Societies. Published by Elsevier B.V. All rights reserved.en_IE
dc.description.sponsorshipThis work was funded by Science Foundation Ireland and developed under Technology Innovation Development (TIDA) Feasibility study (11/TIDA/B2033).en_IE
dc.formatapplication/pdfen_IE
dc.language.isoenen_IE
dc.publisherElsevieren_IE
dc.relation.ispartofImmunology Lettersen
dc.subjectMesenchymal stromal cellsen_IE
dc.subjectMonocytesen_IE
dc.subjectTNF-alphaen_IE
dc.subjectImmunomodulatory potencyen_IE
dc.subjectFlow cytometryen_IE
dc.subjectWhole blood assayen_IE
dc.subjectStem cellsen_IE
dc.subjectDendritic cellsen_IE
dc.subjectMonocytesen_IE
dc.subjectImmunosuppressionen_IE
dc.subjectDiseasesen_IE
dc.subjectTissueen_IE
dc.subjectBlooden_IE
dc.subjectRegenerative medicineen_IE
dc.titleDevelopment of a flow cytometry-based potency assay for measuring the in vitro immunomodulatory properties of mesenchymal stromal cellsen_IE
dc.typeArticleen_IE
dc.date.updated2017-02-03T20:53:56Z
dc.identifier.doi10.1016/j.imlet.2016.07.010
dc.local.publishedsourcehttp://dx.doi.org/10.1016/j.imlet.2016.07.010en_IE
dc.description.peer-reviewedpeer-reviewed
dc.contributor.funder|~|
dc.description.embargo2017-07-20
dc.internal.rssid11421830
dc.local.contactRhodri Ceredig, Room 217, Orbsen Building, Nui Galway. 5916 Email: rhodri.ceredig@nuigalway.ie
dc.local.copyrightcheckedYes
dc.local.versionACCEPTED
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