Thromb Haemost 1968; 20(03/04): 384-396
DOI: 10.1055/s-0038-1651281
Originalarbeiten – Original Articles – Travaux Originaux
Schattauer GmbH

In Vitro Assay of Platelet Adhesiveness with Washed and Tanned Human Red Cells

G Zbinden
1   Department of Medicine, University of Cambridge, Medical School, Cambridge
,
S Tomlin
1   Department of Medicine, University of Cambridge, Medical School, Cambridge
› Author Affiliations
Further Information

Publication History

Publication Date:
27 June 2018 (online)

Summary

An in vitro system is described in which adhesion of blood platelets to washed and tannic acid-treated red cells was assayed quantitatively by microscopic observation. ADP, epinephrine and TAME produced a reversible increase in platelet adhesiveness which was antagonized by AMP. With Evans blue, polyanetholsulfonate, phthalanilide NSC 38280, thrombin and heparin at concentrations above 1-4 u/ml the increase was irreversible. The ADP-induced increase in adhesiveness was inhibited by sodium citrate, EDTA, AMP, ATP and N-ethylmaleimide. EDTA, AMP and the SH-blocker N-ethylmaleimide also reduced spontaneous platelet adhesion to red cells. No significant effects were observed with adenosine, phenprocoumon, 5-HT, phthalanilide NSC 57155, various estrogens, progestogens and fatty acids, acetylsalicylic acid and similarly acting agents, hydroxylamine, glucose and KCN. The method may be useful for the screening of thrombogenic and antithrombotic properties of drugs.

 
  • References

  • 1 Baumgartner H.R, Studer A, Reber K. Influence of 5-hydroxytryptamine, 5-hydroxy-tryptophan, dopamine, norepinephrine and reserpine on thrombotic deposits in the rabbit. Thrombos. Diathes. haemorrh. (Stuttg.) 1964; 12: 168-178
  • 2 Bennett P.N. Effect of glucose on platelet adhesiveness. J. Atheroscl. Res 1967; 7: 53-58
  • 3 Berman H.J, Fulton G.P, Lutz B.R, Pierce D.L. Susceptibility to thrombosis in normal young, aging, cortisone-treated, heparinized and x-irradiated hamsters as tested by topical application of thrombin. Blood 1955; 10: 831-840
  • 4 Booyse F.M, Hoveke T.P, Rafelson Jr. M.E. Density gradient and properties of human platelet populations. Fed. Proc 1968; 27: 569
  • 5 Born G.V.R. Aggregation of blood platelets by adenosine diphosphate and its reversal. Nature (Lond.) 1962; 194: 927-929
  • 6 Born G.V.R, Cross M.J. The aggregation of blood platelets. J. Physiol. (Lond.) 1951; 168: 178-963
  • 7 Bridges J.M, Dolby A.M, Millar J.D.H, Weaver J.A. An effect of D-glucose on platelet stickiness. Lancet 1965; I: 75-77
  • 8 Brinkhous K.M, Read M.S, Mason R.G. Plasma thrombocyte-agglutinating activity and fibrinogen. Lab. Invest 1965; 14: 335-342
  • 9 Constantine J.W, Hochstein F.A. Aggregation of rabbit and guinea-pig platelets by oxime esters. Nature (Lond.) 1966; 210: 164-167
  • 10 Cunnigham G.M, McNicol G.P, Douglas A.S. Effect of anticoagulant drugs on platelet aggregation in the Chandler's tube. Lancet 1965; I: 729-730
  • 11 Davis J.W, Wilson S.J, McField J.R. In vitro and in vivo effects of glucose on human platelet aggregation induced by adenosine diphosphate. Blood 1957; 30: 556
  • 12 Eastham R.D. Rapid adhesive platelet count in whole blood. J. clin. Pathol 1964; 17: 45-46
  • 13 Evans G, Nishizawa E.E, Packham M.A, Mustard J.F. The effect of acetylsalicylic acid (aspirin) on platelet function. Blood 1967; 30: 550
  • 14 Evensen S.A, Jeremic M, Hjort P.F. Intravascular coagulation with generalized Shwartz-man reaction induced by a heparin-like anticoagulant (Liquoid). Thrombos. Diathes. haemorrh. (Stuttg.) 1967; 18: 24-39
  • 15 Fulton G.P, Akers R.P, Lutz B.R. White thrombo-embolism and vascular fragility in the hamster cheek pouch after anticoagulants. Blood 1953; 8: 140-152
  • 16 Gaarder A.J, onsen J, Laland S, Hellem A, Owren P.A. Adenosine diphosphate in red cells as a factor in the adhesiveness of human blood platelets. Nature (Lond.) 1961; 192: 531-532
  • 17 Gast L.F. Influence of aspirin on haemostatic parameters. Ann. rheum. Dis 1964; 23: 500-504
  • 18 Gibson J.G, Gregersen M.I. Toxicity of two vital dyes used in plasma volume determination. Amer. J. Physiol 1953; 113: 50
  • 19 Glynn M.F, Movat H.Z, Murphy E.A, Mustard J.F. Study of platelet adhesiveness and aggregation, with latex particles. J. Lab. clin. Med 1965; 65: 179-201
  • 20 Haslam R.J. Role of adenosine diphosphate in the aggregation of human blood-platelets by thrombin and by fatty acids. Nature (Lond.) 1964; 202: 765-768
  • 21 Hellem A.J. The adhesiveness of human platelets in vitro. Scand. J. clin. Lab. Invest 1960; 12: 1-117 Suppl. 51
  • 22 Hovig T. Aggregation of rabbit blood platelets produced in vitro by saline “extract” of tendons. Thrombos. Diathes. haemorrh. (Stuttg.) 1963; 9: 248-263
  • 23 Kerr J.W, MacAulay I, Pirrie R, Bronte-Stewart B. Platelet-aggregation by phospholipids and free fatty acids. Lancet 1965; I: 1296-1299
  • 24 McDonald L, Edgill M. Action of heparin in ischaemic heart disease. Lancet 1961; I: 844-847
  • 25 McLean J.R, Maxwell R.E, Hertler D. Fibrinogen and adenosine diphosphate-induced aggregation of platelets. Nature (Lond.) 1964; 202: 605-606
  • 26 McLean J.R, Maxwell R.E, Hertler D. Antagonism of platelet aggregation by competitive thrombin substrates. Life Sci 1964; 3: 1313-1317
  • 27 Mertz E.T, Owen G.A. Imidazole buffer: its use in blood clotting studies. Proc. Soc. exp. Biol. (N. Y.) 1940; 43: 204-205
  • 28 Mills D.G.B, Roberts G.C.K. Membrane active drugs and the aggregation of human blood platelets. Nature (Lond.) 1967; 213: 35-38
  • 29 Mitchell J.R.A, Sharp A.A. Platelet clumping in vitro. Brit. J. Haemat 1964; 10: 78-93
  • 30 Moolten S.E, Vroman L. The adhesiveness of blood platelets in thromboembolism and haemorrhagic disorders. I. Measurement of platelet adhesiveness by the glass-wool filter. Amer. J. clin. Path 1949; 19: 701-826
  • 31 Murchison L, Fyfe T. Effects of cigarette smoking on serum-lipids, blood-glucose and platelet adhesiveness. Lancet 1966; II: 182-184
  • 32 Mustard J.F, Glynn M.F, Nishizawa E.E, Packham M.A. Platelet-surface interactions: relationship to thrombosis and hemostasis. Fed. Proc 1967; 26: 106-114
  • 33 Nordøy A, Ødegaard A.E. The influence of citrate and heparin on the adhesiveness of rat platelets and human platelets measured in vitro. Scand. J. clin. Lab. Invest 1963; 15: 399-404
  • 34 Nordøy A, Rørvik T.O. Some effects of adrenaline on rat platelets in vitro and in vivo. Scand. J. clin. Lab. Invest 1965; 17: 151-158
  • 35 O’Brien J.R. The adhesiveness of native platelets and its prevention. J. clin. Path 1961; 14: 140-149
  • 36 O’Brien J.R. Platelet aggregation. Part I. Some effects of the adenosine phosphates, thrombin and cocaine upon platelet adhesiveness. J. clin. Path 1962; 15: 446-455
  • 37 O’Brien J.R. The effect of the alkyl tins on platelet aggregation. Thrombos. Diathes. haemorrh. (Stuttg.) 1963; 9: 330-334
  • 38 O’Brien J.R. Some effects of adrenaline and anti-adrenaline compounds on platelets in vitro and in vivo. Nature (Lond.) 1963; 200: 763-764
  • 39 O’Brien J.R. Aspirin platelet aggregation. Lancet 1968; I: 203
  • 40 Packham M.A, Warrior E.S, Glynn M.F, Senyi A.S, Mustard J.F. Alterations of the response of platelets to surface stimuli by pyrazole compounds. J. exp. Med 1967; 126: 171-188
  • 41 Perkins H.A, Osborn J.J, Gerbode F. The effect of heparin on the platelet count in vitro. With particular reference to the collection of blood for extracorporeal circulation. Amer. J. clin. Path 1958; 30: 397-403
  • 42 Quick A.J. Salicylates and bleeding: the aspirin tolerance test. Amer. J. med. Sci 1966; 252: 265-269
  • 43 Salzman E.W, Chambers D.A. Inhibition of ADP-induced platelet aggregation by substituted amino acids. Nature (Lond.) 1964; 204: 698-700
  • 44 Salzman E.W, Chambers D.A. Incorporation by blood platelets of adenosine diphosphate labelled with carbon-14. Nature (Lond.) 1965; 206: 727-728
  • 45 Salzman E.W, Chambers D.A, Neri L.L. Possible mechanism of aggregation of blood platelets by adenosine diphosphate. Nature (Lond.) 1966; 210: 167-169
  • 46 Shermer R.W, Mason R.G, Wagner R.H, Brinkhous K.M. Studies on thrombin-induced platelet agglutination. J. exp. Med 1961; 114: 905-920
  • 47 Shore P.A, Alpers H.S. Platelet damage and release of amines by microgram quantities of certain fatty acids. Fed. Proc 1963; 22: 504
  • 48 Shore P.A, Alpers H.S. Parallel inhibition of spontaneous and antigen-induced platelet histamine release. Amer. J. Physiol 1963; 205: 348-350
  • 49 Silver M.D, Stehbens W.E, Silver M.M. Platelet reaction to adenosine diphosphate in vivo. Nature (Lond.) 1965; 205: 91-92
  • 50 Skoza L, Zucker M.B, Jerushalmy Z, Grant R. Kinetic studies of platelet aggregation induced by adenosine diphosphate and its inhibition by chelating agents, guanidino compounds, and adenosine. Thrombos. Diathes. haemorrh. (Stuttg.) 1967; 18: 713-725
  • 51 Solum N.O, Stormorken H. Influence of fibrinogen on the aggregation of washed human blood platelets induced by adenosine diphosphate, thrombin, collagen and adrenaline. Scand. J. clin. Lab. Invest 1965; 17 (Suppl. 84) 170-182
  • 52 Spaet T.H, Zucker M.B. Mechanism of platelet plug formation and role of adenosine diphosphate. Amer. J. Physiol 1964; 206: 1267-1274
  • 53 Spooner M, Meyer O.O. The effect of dicoumarol (3,3’-methylenebis[4-hydroxycourQariii]) on platelet adhesiveness. Amer. J. Physiol 1944; 142: 279-283
  • 54 Stormorken H, Lund-Riise Å, Rorvik T.O. Platelet adhesiveness to glass beads: Methodological investigations using automatic platelet counting. Scand. J. clin. Lab. Invest 1965; 17: 183-194
  • 55 Weiner M, Zeltmacher K, Reich G, Shapiro S. Platelet adhesiveness. Blood 1948; 3: 1275-1282
  • 56 Weiss H.J, Aledort L.M. Impaired platelet/connective-tissue reaction in man after aspirin ingestion. Lancet 1967; II: 495-497
  • 57 Whitaker A.N, McKay D.G. Disseminated intravascular coagulation in rabbits by intravenous infusion of epinephrine. Fed. Proc 1968; 27: 569
  • 58 Wright H.P. The adhesiveness of blood platelets in normal subjects with varying concentrations of anticoagulants. J. Path. Bacteriol 1941; 53: 255-262
  • 59 Wright H.P. The adhesiveness of blood platelets in rabbits treated with dicoumarol. J. Path. Bacteriol 1949; 57: 383-385
  • 60 Yamanaka M, Isobe J, Kono Y. Effect of adenosine diphosphate on adhesiveness of platelets in vitro and in vivo. Nature (Lond.) 1967; 216: 175-176
  • 61 York I.M, Rogers W.I, Kensler C.J. The production of thrombi and emboli in the hamster cheek pouch by phthalanilides and related compounds. J. Pharmacol. exp. Ther 1963; 141: 36-49
  • 62 Zbinden G. Biochemische, funktionelle und morphologische Organveränderungen durch Beeinflussung des 5-Hydroxytryptamin-Stoffwechsels. Ergebn. allg. Pathol. path. Anat 1960; 39: 225-259
  • 63 Zbinden G. Modification of the fatty acid-induced thrombocytopenia by anticoagulants and compounds which inhibit in vitro platelet aggregation. J. Pharmacol. exp. Ther 1968; 159: 163-171
  • 64 Zucker M.B. Platelet agglutination and vasoconstriction as factors in spontaneous hemo-stasis in normal, thrombocytopenic, heparinized and hypoprothrombinemic rats. Amer. J. Physiol 1947; 148: 275-288
  • 65 Zucker M.B, Borrelli J. Platelet clumping produced by connective tissue suspensions and by collagen. Proc. Soc. exp. Biol. (N. Y.) 1962; 109: 779-787
  • 66 Zucker M.B, Peterson J. Inhibition of adenosine diphosphate-induced aggregation and other platelet functions by acetylsalicylic acid ingestion. Proc. Soc. exp. Biol. (N. Y.) 1968; 127: 547-551