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Thromb Haemost 2006; 96(06): 781-788
DOI: 10.1160/TH06-05-0242
Platelets and Blood Cells
Schattauer GmbH

Multiple electrode aggregometry: A new device to measure platelet aggregation in whole blood

Orsolya Tóth
1   Institute for Prevention of Cardiovascular Diseases, University of Munich, Munich, Germany
2   1stDepartment of Medicine, University of Pécs, Pécs, Hungary
,
Andreas Calatzis
3   Department of Transfusion Medicine and Hemostaseology, University of Munich, Munich, Germany
,
Sandra Penz
1   Institute for Prevention of Cardiovascular Diseases, University of Munich, Munich, Germany
,
Hajna Losonczy
2   1stDepartment of Medicine, University of Pécs, Pécs, Hungary
,
Wolfgang Siess
1   Institute for Prevention of Cardiovascular Diseases, University of Munich, Munich, Germany
› Author AffiliationsFinancial support: The study was supported by the August-Lenz foundation (O.T.) and the graduate program GK438 “Vascular Biology in Medicine” Deutsche Forschungsgemeinschaft (S.P.; W.S).
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Publication History

Received 05 May 2006

Accepted after resubmission 09 October 2006

Publication Date:
29 November 2017 (online)

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Summary

Several methods are used to analyse platelet function in whole blood. A new device to measure whole blood platelet aggregation has been developed, called multiple electrode platelet aggregometry (MEA). Our aim was to evaluate MEA in comparison with the single platelet counting (SPC) method for the measurement of platelet aggregation and platelet inhibition by aspirin or apyrase in diluted whole blood. Platelet aggregation induced by different concentrations of ADP, collagen and TRAP-6 and platelet inhibition by apyrase or aspirin were determined in citrateor hirudin-anticoagulated blood by MEA and SPC. MEA indicated that spontaneous platelet aggregation was lower, and stimulated platelet aggregation was higher in hirudin- than citrate-anticoagulated blood. In hirudin-anticoagulated, but not citrate-anticoagulated blood, spontaneous platelet aggregation measured by MEA was inhibited by apyrase. For MEA compared with SPC the dose response-curves of agonist-induced platelet aggregation in citrate- and hirudin-blood showed similar EC50 values for TRAP, and higher EC50 values for ADP (non-significant) and collagen (p<0.05). MEA and the SPC method gave similar results concerning platelet-inhibition by apyrase and aspirin. MEA was more sensitive than SPC to the inhibitory effect of aspirin in collagen-induced aggregation. In conclusion, MEA is an easy, reproducible and sensitive method for measuring spontaneous and stimulated platelet aggregation, and evaluating antiplatelet drugs in diluted whole blood. The use of hirudin as an anticoagulant is preferable to the use of citrate. MEA is a promising technique for experimental and clinical applications.

Keywords

Anticoagulation - multiple electrode aggregometry - platelet aggregation - single platelet counting
 

  • References

  • 1 Born GV, Cross MJ. Effect of adenosine diphosphate on the concentration of platelets in circulating blood. Nature 1963; 197: 974-6.
    CrossrefPubMedGoogle Scholar
  • 2 Bartlett GR. Metabolism by man of intravenously administered adenine. Transfusion 1977; 17: 367-73.
    CrossrefPubMedGoogle Scholar
  • 3 Santos MT, Valles J, Marcus AJ. et al. Enhancement of platelet reactivity and modulation of eicosanoid production by intact erythrocytes. A new approach to platelet activation and recruitment. J Clin Invest 1991; 87: 571-80.
    CrossrefPubMedGoogle Scholar
  • 4 Persidsky MD, Ling NS. Separation of platelet-rich plasma by modified centrifugal elutriation. J Clin Apher 1982; 01: 18-24.
    CrossrefPubMedGoogle Scholar
  • 5 Reiss RF, Katz AJ. Optimizing recovery of platelets in platelet rich plasma by the simplex strategy. Transfusion 1976; 16: 370-4.
    PubMedGoogle Scholar
  • 6 Hill RJ, Stenberg PE, Sullam PM. et al. Use of arabinogalactan to obtain washed murine platelets free of contaminating plasma proteins and appropriate for studies of function, morphology, and thrombopoiesis. J Lab Clin Med 1988; 111: 73-83.
    PubMedGoogle Scholar
  • 7 Dyszkiewicz-Korpanty AM, Frenkel EP, Sarode R. Approach to the assessment of platelet function: comparison between optical-based platelet-rich plasma and impedance-based whole blood platelet aggregation methods. Clin Appl Thromb Hemost 2005; 11: 25-35.
    CrossrefPubMedGoogle Scholar
  • 8 Fox SC, Burgess-Wilson M, Heptinstall S. et al. Platelet aggregation in whole blood determined using the Ultra-Flo 100 Platelet Counter. Thromb Haemost 1982; 48: 327-9.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 9 Mandal S, Sarode R, Dash S. et al. Hyperaggregation of platelets detected by whole blood platelet aggregometry in newly diagnosed noninsulin-dependent diabetes mellitus. Am J Clin Pathol 1993; 100: 103-7.
    CrossrefPubMedGoogle Scholar
  • 10 Saniabadi AR, Lowe GD, Forbes CD. et al. Platelet aggregation studies in whole human blood. Thromb Res 1983; 30: 625-32.
    CrossrefPubMedGoogle Scholar
  • 11 Gray RP, Hendra TJ, Patterson DL. et al. “Spontaneous” platelet aggregation in whole blood in diabetic and non diabetic survivors of acute myocardial infarction. Thromb Haemost 1993; 70: 932-6.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 12 Cho NH, Becker D, Dorman JS. et al. Spontaneous whole blood platelet aggregation in insulin-dependent diabetes mellitus: an evaluation in an epidemiologic study. Thromb Haemost 1989; 61: 127-30.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 13 Haseruck N, Erl W, Pandey D. et al. The plaque lipid lysophosphatidic acid stimulates platelet activation and platelet-monocyte aggregate formation in whole blood: involvement of P2Y1 and P2Y12 receptors. Blood 2004; 103: 2585-92.
    CrossrefPubMedGoogle Scholar
  • 14 Cardinal DC, Flower RJ. The electronic aggregometer: a novel device for assessing platelet behavior in blood. J Pharmacol Methods 1980; 03: 135-58.
    CrossrefPubMedGoogle Scholar
  • 15 Riess H, Braun G, Brehm G. et al. Critical evaluation of platelet aggregation in whole human blood. Am J Clin Pathol 1986; 85: 50-6.
    CrossrefPubMedGoogle Scholar
  • 16 Ingerman-Wojenski C, Smith JB, Silver MJ. Evaluation of electrical aggregometry: comparison with optical aggregometry, secretion of ATP, and accumulation of radiolabeled platelets. J Lab Clin Med 1983; 101: 44-52.
    PubMedGoogle Scholar
  • 17 Wallen NH, Ladjevardi M, Albert J. et al. Influence of different anticoagulants on platelet aggregation in whole blood; a comparison between citrate, low molecular mass heparin and hirudin. Thromb Res 1997; 87: 151-7.
    CrossrefPubMedGoogle Scholar
  • 18 Phillips DR, Teng W, Arfsten A. et al. Effect of Ca2+ on GP IIb-IIIa interactions with integrilin: enhanced GP IIb-IIIa binding and inhibition of platelet aggregation by reductions in the concentration of ionized calcium in plasma anticoagulated with citrate. Circulation 1997; 96: 1488-94.
    CrossrefPubMedGoogle Scholar
  • 19 Storey RF, Wilcox RG, Heptinstall S. Differential effects of glycoprotein IIb/IIIa antagonists on platelet microaggregate and macroaggregate formation and effect of anticoagulant on antagonist potency. Implications for assay methodology and comparison of different antagonists. Circulation 1998; 98: 1616-21.
    CrossrefPubMedGoogle Scholar
  • 20 Sharp DS, Beswick AD, O’Brien JR. et al. The association of platelet and red cell count with platelet impedance changes in whole blood and light-scattering changes in platelet rich plasma: evidence from the Caerphilly Collaborative Heart Disease Study. Thromb Haemost 1990; 64: 211-5.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 21 Armstrong R, May JA, Losche W. et al. Factors that contribute to spontaneous platelet aggregation and streptokinase-induced aggregation in whole blood. Thromb Haemost 1995; 73: 297-303.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 22 Sathiropas P, Marbet GA, Sahaphong S. et al. Detection of small inhibitory effects of acetylsalicylic acid (ASA) by platelet impedance aggregometry in whole blood. Thromb Res 1988; 51: 55-62.
    CrossrefPubMedGoogle Scholar
  • 23 Storey RF, Newby LJ, Heptinstall S. Effects of P2Y(1) and P2Y(12) receptor antagonists on platelet aggregation induced by different agonists in human whole blood. Platelets 2001; 12: 443-7.
    CrossrefPubMedGoogle Scholar