Thromb Haemost 1977; 37(02): 291-308
DOI: 10.1055/s-0038-1649230
Original Article
Schattauer GmbH

Properties of Washed Human Platelets

Raelene L Kinlough-Rathbone
1   Department of Pathology, McMaster University, Hamilton
2   Department of Biochemistry, University of Toronto, Toronto, Canada
,
J Fraser Mustard
1   Department of Pathology, McMaster University, Hamilton
2   Department of Biochemistry, University of Toronto, Toronto, Canada
,
Marian A Packham
1   Department of Pathology, McMaster University, Hamilton
2   Department of Biochemistry, University of Toronto, Toronto, Canada
,
Dennis W Perry
1   Department of Pathology, McMaster University, Hamilton
2   Department of Biochemistry, University of Toronto, Toronto, Canada
,
Hans-Joachim Reimers
1   Department of Pathology, McMaster University, Hamilton
2   Department of Biochemistry, University of Toronto, Toronto, Canada
,
Jean-Pierre Cazenave
1   Department of Pathology, McMaster University, Hamilton
2   Department of Biochemistry, University of Toronto, Toronto, Canada
› Author Affiliations
Further Information

Publication History

Received 03 August 1976

Accepted 10 December 1976

Publication Date:
03 July 2018 (online)

Summary

We have shown previously that washed human platelets resuspended in Tyrode solution containing albumin and apyrase maintain their disc shape and their ability to aggregate upon the addition of low concentrations of ADP, providing fibrinogen is added to the suspending medium. We have now examined their responses to other aggregating and release-inducing agents. Collagen, arachidonate, thrombin, immune serum globulin, the ionophore A 23, 187 and phytohaemagglutinin from Phaseolus vulgaris caused aggregation and release of granule contents. The response to adrenaline was variable. Serotonin caused the platelets to change shape but no aggregation or release occurred. Addition of a small amount of plasma was necessary for ristocetin-induced aggregation. Polylysine caused immediate platelet-to-platelet adherence with little or no release of granule contents. Responses to collagen or thrombin were greater in a modified medium containing magnesium but no calcium; in this medium, aggregation caused by ADP or polylysine was followed by the release of granule contents whereas these agents caused aggregation without release in a medium with both calcium and magnesium. When protein was omitted from the suspending medium, platelet aggregation in response to ADP was variable. In this medium, collagen and thrombin caused more extensive release than in the albumin-containing medium. Aggregation by polylysine was accompanied by release and extensive lysis in the protein-free medium. Thus, the composition of the final resuspending medium has a major effect on the responses of washed human platelets to aggregating agents.

 
  • References

  • 1 Ardlie N. G, Packham M. A, Mustard J. F. Adenosine diphosphate-induced platelet aggregation in suspensions of washed rabbit platelets. British Journal of Haematology 1970; 19: 7
  • 2 Ardlie N. G, Perry D. W, Packham M. A, Mustard J. F. Influence of apyrase on stability of suspensions of washed rabbit platelets. Proceedings of the Society for Experimental Biology and Medicine 1971; 136: 1021
  • 3 Aster R. H, Jandl J. H. Platelet sequestration in man. I. Methods. Journal of Laboratory and Clinical Investigation 1964; 43: 843
  • 4 Bang N. U, Hansen M. S, Heidenreich R. O. The influence of plasma proteins on platelet function and metabolism. In Platelets, Recent Advances in Basic Research and Clinical Aspects. Exerpta Medica International Congress Series No. 357, Exerpta Medica. Amsterdam: 1975. p. 118
  • 5 Bang N. U, Heidenreich R. O, Trygstad C. W. Plasma protein requirement for human platelet aggregation. In Platelets and Their Role in Hemostasis. Annals of the New York Academy of Sciences 1972; 201: 280
  • 6 Bergmeyer H. U, Bernt E, Hess B. Lactic dehydrogenase. In Bergmeyer H. U. (ed.) Methods in Enzymatic Analysis. Academic Press; New York: 1965. p. 736
  • 7 Born G. V. R, Cross M. J. The aggregation of blood platelets. Journal of Physiology (Lond) 1963; 168: 178
  • 8 Born G. V. R, Gillson R. E. Studies on the uptake of 5-hydroxytryptamine by blood platelets. Journal of Physiology (Lond) 1959; 146: 472
  • 9 Born G. V. R, Michal F. 5-hydroxytryptamine receptors of platelets. In Biochemistry and Pharmacology of Platelets. Ciba Foundation Symposium, 35 (new series). Elsevier/Exerpta Medica/North Holland; Amsterdam.: p. 287 1975
  • 10 Cazenave J.-P, Packham M. A, Guccione M. A, Mustard J. F. Inhibition of platelet adherence to damaged surface of rabbit aorta. Journal of Laboratory and Clinical Medicine 1975; 86: 551
  • 11 Cazenave J.-P, Packham M. A, Mustard J. F. Adherence of platelets to a collagen-coated surface: development of a quantitative method. Journal of Laboratory and Clinical Medicine 1973; 82: 978
  • 12 Cross M. J. Effect of fibrinogen on the aggregation of platelets by adenosine diphosphate. Thrombosis et Diathesis Haemorrhagica 1964; 12: 524
  • 13 Day H. J, Holmsen H, Zucker M. B. Methods for separating platelets from blood and plasma. Thrombosis et Diathesis Haemorrhagica 1975; 33: 648
  • 14 Doery J. C. G, Hirsh J, Mustard J. F. Energy metabolism in washed human platelets responsive to ADP: comparison with platelets in plasma. British Journal of Haematology 1973; 25: 657
  • 15 Goldsmith H. L, Yu S. S. K, Marlow J. Fluid mechanical stress and the platelet. Thrombosis et Diathesis Haemorrhagica 1975; 34: 32
  • 16 Greenberg J, Packham M. A, Cazenave J.-P, Reimers H.-J, Mustard J. F. Effects on platelet function of removal of platelet sialic acid by neuraminidase. Laboratory Investigation 1975; 32: 476
  • 17 Holmsen H, Setkowsky C. A, Lages B, Day H. J, Weiss H, Scrutton M. C. Content and thrombin-induced release of acid hydrolases in gel-filtered platelets from patients with storage pool disease. Blood 1975; 46: 131
  • 18 Holmsen H, Storm E, Day H. J. Determination of ATP and ADP in blood platelets: a modification of the firefly luciferase assay for plasma. Analytical Biochemistry 1972; 46: 489
  • 19 Holmsen H, Weiss H. J. Hereditary defect in the platelet release reaction caused by a deficiency in the storage pool of adenine nucleotides. British Journal of Haematology 1970; 19: 643
  • 20 Holmsen I, Holmsen H. Partial purification and characterization of an ADP phosphohydrolase from human plasma. Thrombosis et Diathesis Haemorrhagica 1971; 26: 177
  • 21 Howard M, Sawers R. J, Firkin B. G. Ristocetin- a means of differentiating von Wille-brand’s disease into two groups. Blood 1973; 41: 687
  • 22 Hutton R. A, Howard M, Deykin D, Hardisty R. M. Methods for the separation of platelets from plasma. A comparison of functional and morphological integrity. Thrombosis et Diathesis Haemorrhagica 1974; 31: 119
  • 23 Jenkins C. S. P, Meyer D, Dreyfus M. D, Larrieu M.-J. Willebrand factor and ristocetin. I. Mechanism of ristocetin-induced platelet aggregation. British Journal of Haematology 1974; 28: 561
  • 24 Jenkins C. S. P, Packham M. A, Guccione M. A, Mustard J. F. Modification of platelet adherence to protein-coated surfaces. Journal of Laboratory and Clinical Medicine 1973; 81: 280
  • 25 Joist J. H, Dolezel G, Lloyd J. V, Kinlough-Rathbone R. L, Mustard J. F. Platelet factor-3 availability and the platelet release reaction. Journal of Laboratory and Clinical Medicine 1974; 84: 474
  • 26 Kinlough-Rathbone R. L, Chahil A, Packham M. A, Reimers H.-J, Mustard J. F. Effect of ionophore A 23 187 on thrombin-degranulated washed rabbit platelets. Thrombosis Research 1975; 7: 435
  • 27 Kinlough-Rathbone R. L, Packham M. A, Mustard J. F. The effect of glucose on adenosine diphosphate-induced platelet aggregation. Journal of Laboratory and Clinical Medicine 1970; 75: 780
  • 28 Kinlough-Rathbone R. L, Reimers H.-J, Mustard J. F, Packham M. A. Sodium arachidonate can induce platelet shape change and aggregation which are independent of the release reaction. Science 1976; 192: 1011
  • 29 Lages B, Scrutton M. C, Holmsen H. Studies on gel-filtered human platelets: isolation and characterization in a medium containing no added Ca2+, Mg2+, or K+ . Journal of Laboratory and Clinical Medicine 1975; a 85: 811
  • 30 Lages B, Scrutton M. C, Holmsen H, Day H. J, Weiss H. J. Metal ion contents of gel-filtered platelets from patients with storage pool disease. Blood 1975; b 46: 119
  • 31 Levy-Toledano S, Rendu F, Besson P, Caen J. Aggregation of human gel-filtered platelets: requirement of apyrase and proteins. Revue Européenne d’Etudes Cliniques et Biologiques 1972; 17: 513
  • 32 Lyman D. J, Brash J. L, Chaikin S. W, Klein K. G, Carini M. The effect of chemical structure and surface properties of synthetic polymers on coagulation of blood. II. Protein and platelet interaction with polymer surfaces. Transactions: American Society for Artificial Internal Organs 1968; 14: 250
  • 33 Mason R. G, Read M. S, Shermer R. W. Comparison of certain functions of human platelets separated from blood by various means. American Journal of Pathology 1974; 76: 323
  • 34 Mills D. C. B. Changes in the adenylate energy charge in blood platelets induced by adenosine diphosphate. Nature New Biology 1973; 243: 220
  • 35 Mills D. C. B, Thomas D. Blood platelet nucleotides in man and other species. Nature 1969; 222: 991
  • 36 Mohammad S. F, Reddick R. L, Mason R. G. Characterization of human platelets separated from blood by ADP-induced aggregation. American Journal of Pathology 1975; 79: 81
  • 37 Molnar J, Lorand L. Studies on apyrases. Archives of Biochemistry and Biophysics 1961; 93: 353
  • 38 Mustard J. F, Packham M. A. Factors influencing platelet function: adhesion release and aggregation. Pharmacological Reviews 1970; 22: 97
  • 39 Mustard J. F, Perry D. W, Ardlie N. G, Packham M. A. Preparation of suspensions of washed platelets from humans. British Journal of Haematology 1972; 22: 193
  • 40 Mustard J. F, Perry D. W, Kinlough-Rathbone R. L, Packham M. A. Factors responsible for ADP-induced release reaction of human platelets. American Journal of Physiology 1975; 228: 1757
  • 41 Nicholls D. G, Hampton J. R. Density gradient separation of human platelets from plasma and the role of plasma in adenosine diphosphate induced platelet electrophoretic mobility change. Thrombosis et Diathesis Haemorrhagica 1972; 27: 425
  • 42 O’Brien J. R. Changes in platelet membranes possibly associated with platelet stickiness. Nature 1966; 212: 1057
  • 43 O’Brien J. R, Heywood J. B. Effect of aggregating agents and their inhibitors on the mean platelet shape. Journal of Clinical Pathology 1966; 19: 148
  • 44 Olson J. D, Fass D.N, Bowie E.J. W, Mann K. G. Ristocetin-induced aggregation of gel filtered platelets. A study of von Willebrand’s disease and the effect of aspirin. Thrombosis Research 1973; 3: 501
  • 45 Packham M. A, Ardlie N. G, Mustard J. F. Effect of adenine compounds on platelet aggregation. American Journal of Physiology 1969; a 217: 1009
  • 46 Packham M. A, Evans G, Glynn M. F, Mustard J. F. The effect of plasma proteins on the interaction of platelets with glass surfaces. Journal of Laboratory and Clinical Medicine 1969; b 4: 868
  • 47 Packham M. A, Guccione M. A, Chang P.-L, Mustard J. F. Platelet aggregation and release: effects of low concentrations of thrombin or collagen. American Journal of Physiology 1973; 225: 38
  • 48 Packham M. A, Warrior E. S, Glynn M. F, Senyi A. S, Mustard J. F. Alteration of the response of platelets to surface stimuli by pyrazole compounds. Journal of Experimental Medicine 1967; 126: 171
  • 49 Reimers H.-J, Packham M. A, Mustard J. F. Labeling of the releasable adenine nucleotides of washed human platelets. Blood 1977; 49: 89
  • 50 Rossi E. The effect of albumin upon the loss of enzymes from washed platelets. Journal of Laboratory and Clinical Medicine 1972; 79: 240
  • 51 Rozenberg M. C, Holmsen H. Adenine nucleotide metabolism of blood platelets. IV. Platelet aggregation response to exogenous ATP and ADP. Biochimica et Biophysica Acta 1968; 157: 280
  • 52 Salzman E. W. Role of platelets in blood-surface interactions. Federation Proceedings 1971; 30: 1503
  • 53 Sheppard G, Marlow C. G. The simultaneous measurement of 51Cr and 14C by liquid scintillation counting. International Journal of Applied Radiation and Isotopes 1971; 22: 125
  • 54 Tangen O, Andrae M.-L, Nilsson B. E. Nucleotide leakage from platelets in artificial media: prevention by albumin and other macromolecules and relation to ADP-induced platelet aggregation. Scandinavian Journal of Haematology 1973; 11: 241
  • 55 Tangen O, Berman H. J, Marfey P. Gel filtration: a new technique for separation of platelets from plasma. Thrombosis et Diathesis Haemorrhagica 1971; 25: 268
  • 56 Walsh P. N. Albumin density gradient separation and washing of platelets and the study of platelet coagulant activities. British Journal of Haematology 1972; 22: 205
  • 57 Weiss H. J, Rogers J, Brand H. Defective ristocetin-induced platelet aggregation in von Willebrand’s disease and its correction by factor VIII. Journal of Clinical Investigation 1973; 52: 2697