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Platelet function testing at low platelet counts: When can you trust your analysis?
Department of Haematology and Department of Clinical and Experimental Medicine, Linköping University, Linköping, Sweden; Australian Centre for Blood Diseases, Monash University, Melbourne, Australia.
Department of Clinical and Experimental Medicine, Linköping University, Linköping, Sweden.
Department of Clinical and Experimental Medicine, Linköping University, Linköping, Sweden.
Örebro University, School of Medical Sciences. Department of Clinical Chemistry and Department of Clinical and Experimental Medicine, Linköping University, Linköping, Sweden; Cardiovascular Research Centre, Örebro University, School of Medical Sciences, Örebro, Sweden.ORCID iD: 0000-0002-1920-3962
2019 (English)In: Research and Practice in Thrombosis and Haemostasis, E-ISSN 2475-0379, Vol. 3, no 2, p. 285-290Article in journal (Refereed) Published
Abstract [en]

Background: Although flow cytometry is often brought forward as a preferable method in the setting of thrombocytopenia, the relative effects of low sample counts on results from flow cytometry-based platelet function testing (FC-PFT) in comparison with light transmission aggregometry (LTA) and multiple electrode aggregometry (MEA) has not been reported.

Objectives: To compare the effects of different sample platelet counts (10, 50, 100, and 200x10(9)L(-1)) on platelet activation measured with FC-PFT, LTA, and MEA using the same anticoagulant and agonist concentrations as for the commercial MEA test.

Methods: Platelets were stimulated with two commonly used platelet agonists (ADP [6.5 mu molL(-1)] and PAR1-AP [TRAP, 32 mu molL(-1)]). The specified sample platelet counts were obtained by combining platelet-rich and platelet poor hirudinized plasma in different proportions with or without red blood cells.

Results: For FC, P-selectin exposure and PAC-1 binding was reduced at 10x10(9)L(-1) after stimulation with PAR1-AP (by approximately 20% and 50%, respectively), but remained relatively unchanged when ADP was used as agonist (n=9). The platelet count-dependent effects observed with PAR1-AP were eliminated when samples were pre-incubated with apyrase, implying that reduced purinergic signaling was the main underlying factor (n=5). Both aggregometry-based PFTs showed a 50% reduction at 50x10(9)L(-1) and more than 80% reduction at 10x10(9)L(-1), irrespective of agonist used (n=7).

Conclusions: Although FC-PFT is generally preferable to aggregometry-based PFTs in situations with low sample platelet counts, a careful optimization of experimental parameters is still required in order to eliminate platelet count-related effects.

Place, publisher, year, edition, pages
Wiley-Blackwell, 2019. Vol. 3, no 2, p. 285-290
Keywords [en]
platelet activation, platelet aggregation, platelet count, platelet function tests, thrombocytopenia
National Category
Hematology
Identifiers
URN: urn:nbn:se:oru:diva-73960DOI: 10.1002/rth2.12193ISI: 000464657400021PubMedID: 31011713OAI: oai:DiVA.org:oru-73960DiVA, id: diva2:1307755
Funder
Swedish Heart Lung Foundation, 2017-0318Swedish Society of Medicine, SLS-787211
Note

Funding Agencies:

Linköpings Universitet

Region Östergötland

LiU Fund of U; LiU Research Fellows Programme

Available from: 2019-04-29 Created: 2019-04-29 Last updated: 2019-04-29Bibliographically approved

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Ramström, Sofia

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