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Blood. 2015 Sep 3;126(10):e11-8. doi: 10.1182/blood-2015-01-621656. Epub 2015 Jul 27.

Novel whole blood assay for phenotyping platelet reactivity in mice identifies ICAM-1 as a mediator of platelet-monocyte interaction.

Author information

1
William Harvey Research Institute, Barts & the London School of Medicine & Dentistry, Queen Mary University of London, London, United Kingdom;
2
William Harvey Research Institute, Barts & the London School of Medicine & Dentistry, Queen Mary University of London, London, United Kingdom; National Heart & Lung Institute, Imperial College London, London, United Kingdom; and.
3
Leukocyte Adhesion Laboratory, Cancer Research United Kingdom, London Research Institute, London, United Kingdom.

Abstract

Testing of platelet function is central to the cardiovascular phenotyping of genetically modified mice. Traditional platelet function tests have been developed primarily for testing human samples and the volumes required make them highly unsuitable for the testing of mouse platelets. This limits research in this area. To address this problem, we have developed a miniaturized whole blood aggregometry assay, based on a readily accessible 96-well plate format coupled with quantification of single platelet depletion by flow cytometric analysis. Using this approach, we observed a concentration-dependent loss of single platelets in blood exposed to arachidonic acid, collagen, U46619 or protease activated receptor 4 activating peptide. This loss was sensitive to well-established antiplatelet agents and genetic manipulation of platelet activation pathways. Observations were more deeply analyzed by flow cytometric imaging, confocal imaging, and measurement of platelet releasates. Phenotypic analysis of the reactivity of platelets taken from mice lacking intercellular adhesion molecule (ICAM)-1 identified a marked decrease in fibrinogen-dependent platelet-monocyte interactions, especially under inflammatory conditions. Such findings exemplify the value of screening platelet phenotypes of genetically modified mice and shed further light upon the roles and interactions of platelets in inflammation.

PMID:
26215112
PMCID:
PMC4559940
DOI:
10.1182/blood-2015-01-621656
[Indexed for MEDLINE]
Free PMC Article

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