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Generation and characterisation of equine bone marrow-derived macrophages

Research output: Contribution to conferencePoster

Original languageEnglish
Publication statusPublished - 5 Sep 2018
EventEuropean Veterinary Immunology Workshop - Utrecht, Netherlands
Duration: 5 Sep 20187 Sep 2018

Conference

ConferenceEuropean Veterinary Immunology Workshop
CountryNetherlands
CityUtrecht
Period5/09/187/09/18

Abstract

Introduction: Bone marrow-derived macrophages (BMDMs) are primary macrophage cells and are used in vitro as a model for tissue resident macrophages of the innate immune system.
Material and methods: A protocol previously described by Kapetanovic et al, was adapted [1]. Bone marrow from ribs of adult horses was isolated and cultured in human macrophage colony-stimulating factor (rhCSF-1) for 7-10 days. Once differentiated, cells were characterised phenotypically and functionally using microscopy, flow cytometry and phagocytosis assays.
Results: Cell populations showed the expected stellate, adherent morphology of macrophages. Differentiated BMDMs phagocytosed Zymosan and pHrodo Red BioParticles showing that these cells are functional macrophages. Cells expressed surface CD14 but did not express CD163. Following stimulation with LPS, cells did not metabolise arginine to produce nitric oxide (unlike mouse), but metabolised tryptophan through the induction of indoleamine 2,3-dioxygenase, similar to pig and human BMDMs.
Conclusion: A protocol was optimised to produce populations of equine bone-marrow-derived macrophages (eqBMDMs) by cultivation of bone marrow in rhCSF1. Morphological features, the ability to phagocytose particles and the ability to produce cytokines in response to LPS confirmed the cells were macrophages. The method described allows generation of a homogenous population of eqBMDMs to further study their role in the equine innate immune system.
References: 1. Kapetanovic R, Fairbairn L, Beraldi D, Sester D, Archibald A, Tuggle C and Hume D. Pig bone marrow-derived macrophages resemble human macrophages in their response to bacterial lipopolysaccharide. J Immunol. 2012;188:3382-3394

Event

European Veterinary Immunology Workshop

5/09/187/09/18

Utrecht, Netherlands

Event: Conference

ID: 69967290