Thromb Haemost 1976; 36(01): 208-220
DOI: 10.1055/s-0038-1648026
Original Article
Schattauer GmbH

The Use of a Chelating Ion-Exchange Resin to Evaluate the Effects of the Extracellular Calcium Concentration on Adenosine Diphosphate Induced Aggregation of Human Blood Platelets

Stanley Heptinstall
1   University Department of Medicine, General Hospital, Nottingham, NG1 6HA, England.
› Author Affiliations
Further Information

Publication History

Received 10 December 1975

Accepted 20 March 1976

Publication Date:
03 July 2018 (online)

Summary

1. An ion-exchange resin, Chelex 100, has been used to prepare suspensions of human blood platelets in calcium and magnesium depleted plasma.

2. Extracellular calcium is required for platelet aggregation when induced by adenosine diphosphate (ADP). Magnesium only supports aggregation provided that a small amount of calcium is present in the plasma.

3. The extent of platelet aggregation depends upon the concentration of calcium in the plasma. There is an optimum concentration of calcium with which the maximum amount of aggregation is obtained in response to any single concentration of ADP. This optimum calcium concentration is below the physiological level. Higher calcium concentrations reduce the extent of aggregation by enhancing the rate of disaggregation and high magnesium concentrations have the same effect. It is possible that free ADP levels are reduced as a result of ADP-divalent cation complex formation.

4. Platelets were found to contain 18.6 (S.D. ±1.1) × 10–6 mol Ca and 9.3 (S.D. ±1.0) × 10–6 mol Mg per 1011 cells.

 
  • References

  • 1 Adams J, Heptinstall S, Mitchell J. R. A. 1975; A six-channel automated platelet aggregometer. Thrombosis et Diathesis Haemorrhagica 34: 821.
  • 2 Aledort L. M, Hirsch C, Udkow M, Puskin E. 1969; Effect of anticoagulants on calcium and magnesium content of human platelets. Transfusion 9: 212.
  • 3 Ardlie N. G. 1968; ADP-induced platelet aggregation without a plasma cofactor. Federation Proceedings 27: 321.
  • 4 Bio-Rad Laboratories. 1973 Separating metals using Chelex 100 chelating resin. Technical Bulletin 114..
  • 5 Born G. V. R. 1962; Aggregation of blood platelets by adenosine diphosphate and its reversal. Nature 194: 927.
  • 6 Born G. V. R. 1970; Observations on the change in shape of blood platelets brought about by adenosine diphosphate. Journal of Physiology 209: 487.
  • 7 Born G. V. R, Cross M. J. 1963; The aggregation of blood platelets. Journal of Physiology 168: 178.
  • 8 Born G. V. R, Cross M. J. 1964; Effects of inorganic ions and of plasma proteins on the aggregation of blood platelets by adenosine diphosphate. Journal of Physiology 170: 397.
  • 9 Cousin C, Caen J. 1964; Dosage du magnesium et du calcium dans les plaquettes sanguines humaines. Revue Francais Etudes Cliniques et Biologiques IX: 520.
  • 10 Detwiler T. C, Feinman R. D. 1973; Kinetics of the thrombin-induced release of calcium (II) by platelets. Biochemistry 12: 282.
  • 11 Eagon R. G. 1969; Cell wall-associated inorganic substances from Pseudomonas aeruginosa. Canadian Journal of Microbiology 15: 235.
  • 12 Han P, Ardlie N. G. 1974; Heparin, platelets and blood coagulation: Implications for low dose heparin prophylactic regimens in venous thrombosis. British Journal of Haematology 27: 253.
  • 13 Haslam R. J. 1964; Role of adenosine diphosphate in the aggregation of human blood platelets by thrombin and by fatty acids. Nature 202: 765.
  • 14 Hovig T. 1964; The effect of calcium and magnesium on rabbit blood platelet aggregation in vitro . Thrombosis et Diathesis Haemorrhagica 12: 179.
  • 15 Kinlough-Rathbone R. L, Perry D. W, Mustard J. F. 1974; The role of calcium and magnesium in the reaction of ADP with platelets. Federation Proceedings 33: 611.
  • 16 Lages B, Scrutton M. C, Holmsen H. 1975; Studies on gel-filtered human platelets: isolation and characterisation in a medium containing no added Ca++, Mg++, or K+ . Journal of Laboratory and Clinical Medicine 85: 811.
  • 17 MacFarlane D. E, Walsh P. N, Mills D. C. B, Holmsen H, Day H. G. 1975; The role of thrombin in ADP-induced platelet aggregation and release: a critical evaluation. British Journal of Haematology 30: 457.
  • 18 MacMillan D. C, Oliver J. F. 1965; The initial changes in platelet morphology following the addition of adenosine diphosphate. Journal of Atherosclerosis Research 5: 440.
  • 19 Maupin B. 1953; Acquisitions recentes sur l’histochemie et la biochemie des plaquettes sanguines de l’homme. Revue d’Hématologie 8: 302.
  • 20 Mills D. C. B, Robb I. A, Roberts G. C. K. 1968; The release of nucleotides, 5-hydroxytrypt-amine and enzymes from human blood platelets during aggregation. Journal of Physiology 195: 715.
  • 21 Mitchell J. R. A, Sharp A. A. 1964; Platelet clumping in vitro . British Journal of Haematology 10: 78.
  • 22 Murer E. H, Holme R. 1970; A study of the release of calcium from human blood platelets and its inhibition by metabolic inhibitors, N-ethylmaleimide and aspirin. Biochimica Biophysica Acta 222: 197.
  • 23 O’Brien J. R. 1962; Platelet aggregation. II Some results from a new method of study. Journal of Clinical Pathology 15: 452.
  • 24 Sillen L. G, MARTELL A. E. 1964. Stability constants of metalion complexes. Special Publications 17 (1964) and 25 (1971). The Chemical Society; London.:
  • 25 Skoza L, Zucker M. B, Jerushalmy Z, Grant R. 1967; Kinetic studies of platelet aggregation induced by adenosine diphosphate and its inhibition by chelating agents, guanidino compounds, and adenosine. Thrombosis et Diathesis Haemorrhagica 18: 713.
  • 26 Stewart C. P, Dunlop D. 1962. Clinical Chemistry in Practical Medicine. 6. Livingstone: Edinburgh; 236.
  • 27 Stewart W. K, Hutchinson F, Fleming L. W. 1963; The estimation of magnesium in serum and urine by atomic absorption spectrophotometry. Journal of Laboratory and Clinical Medicine 61: 858.
  • 28 Tangen O, McKinnon E. L, Berman H. J. 1973; On the fine structure and aggregation requirements of gel-filtered platelets. Scandinavian Journal of Haematology 10: 96.
  • 29 Wallach D. F. H, Surgenor D. M, Steele B. B. 1958; Calcium-lipid complexes in human platelets. Blood 13: 589.
  • 30 Zucker M. B, Zaccardi J. B. 1964; Platelet shape change induced by adenosine diphosphate and prevented by adenosine monophosphate. Federation Proceedings 23: 299.