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Development of a Method to Quantify Platelet Adhesion and Aggregation Under Static Conditions

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Abstract

Platelets are important players in hemostasis and thrombosis. Thus, accurate assessment of platelet function is crucial for identifying platelet function disorders and measuring the efficacy of antiplatelet therapies. We have developed a novel platelet aggregation technique that utilizes the physical parameter of platelet concentration in conjunction with volume and mass measurements to evaluate platelet adhesion and aggregation. Platelet aggregates were formed by incubating purified platelets on fibrinogen- or fibrillar collagen-coated surfaces at platelet concentrations ranging from 20,000 to 500,000 platelets/μL. Platelets formed aggregates under static conditions in a platelet concentration-dependent manner, with significantly greater mean volume and mass at higher platelet concentrations (≥400,000 platelets/μL). We show that a platelet glycoprotein IIb/IIIa inhibitor abrogated platelet–platelet aggregation, which significantly reduced the volume and mass of the platelets on the collagen surface. This static platelet aggregation technique is amenable to standardization and represents a useful tool to investigate the mechanism of platelet activation and aggregation under static conditions.

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Abbreviations

BSA:

Bovine serum albumin

DIC:

Differential interference contrast

NA:

Numerical aperture

PBS:

Phosphate buffered saline

GP:

Glycoprotein

HTDIC:

Hilbert-transform differential interference contrast

NIQPM:

Non-interferometric quantitative phase microscopy

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Acknowledgments

This work was supported by the National Institutes of Health (R01HL101972, O.J.T.M; R01HL101972-S1, F.A.C.) and a Medical Research Foundation Early Clinical Investigator Award (K.G.P.). S.M.B. is a Whitaker International Fellow. O.J.T.M. is an American Heart Association Established Investigator (13EIA12630000).

Conflict of interest

S.M.B., F.A.C., K.G.P, and O.J.T.M. declare that they have no conflicts of interest.

Ethical Standards

All human subjects research was carried out in accordance with institutional guidelines approved by the Oregon Health & Science University Institutional Review Board. No animal studies were carried out by the authors for this article.

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Authors and Affiliations

  1. Department of Biomedical Engineering, School of Medicine, Oregon Health & Science University, 3303 SW Bond Ave., Portland, OR, 97239, USA

    Sandra M. Baker-Groberg, Flor A. Cianchetti, Kevin G. Phillips & Owen J. T. McCarty

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  1. Sandra M. Baker-Groberg
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  2. Flor A. Cianchetti
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  3. Kevin G. Phillips
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  4. Owen J. T. McCarty
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Corresponding author

Correspondence to Sandra M. Baker-Groberg.

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Associate Editor Michael R. King oversaw the review of this article.

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Baker-Groberg, S.M., Cianchetti, F.A., Phillips, K.G. et al. Development of a Method to Quantify Platelet Adhesion and Aggregation Under Static Conditions. Cel. Mol. Bioeng. 7, 285–290 (2014). https://doi.org/10.1007/s12195-014-0328-9

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  • DOI: https://doi.org/10.1007/s12195-014-0328-9

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