Thromb Haemost 1972; 27(03): 425-433
DOI: 10.1055/s-0038-1649383
Originalarbeiten – Original Articles – Travaux Originaux
Schattauer GmbH

Density Gradient Separation of Human Platelets from Plasma and the Role of Plasma in Adenosine Diphosphate Induced Platelet Electrophoretic Mobility Changes

D. G Nicholls
1   Department of Medicine, Nottingham University, England
,
J. R Hampton
1   Department of Medicine, Nottingham University, England
› Author Affiliations
Further Information

Publication History

Publication Date:
29 June 2018 (online)

Summary

1. An albumin density gradient separation technique is described for separating human blood platelets from plasma.

2. The platelet resuspensions aggregate on the addition of 10-6 M adenosine diphosphate (ADP) in the presence of calcium and fibrinogen.

3. Platelet resuspension in the presence of plasma show a time dependant recovery of reversible aggregation.

4. In the presence of calcium and fibrinogen platelet resuspensions show ADP- induced electrophoretic mobility changes which are the same as those of platelets not separated from their native plasma.

5. Abnormal electrophoretic sensitivity to ADP in platelets from subjects with a history of occlusive arterial disease is retained on separation from plasma.

6. The electrophoretic response of platelet resuspensions to phosphatidyl choline is similar to that of unwashed platelets. The use of resuspended platelets has shown that calcium and fibrinogen are not required for this response.

 
  • References

  • 1 Hampton J. R, Mitchell J. R. A. Effect of aggregating agents on the electrophoretic mobility of human platelets. Brit. med. J I: 1074 1966;
  • 2 Hampton J. R, Mitchell J. R. A. A transferable factor causing abnormal platelet behaviour in vascular disease. Lancet II: 764 1966;
  • 3 Bolton C. H, Hampton J. R, Mitchell J. R. A. Nature of the transferable factor which causes abnormal platelet behaviour in vascular disease. Lancet II: 1101 1967;
  • 4 Hampton J. R, Bolton C. H. Effect of phospholipids on platelet electrophoretic mobility. J. atheroscler. Res 09: 131 1969;
  • 5 Ardlie N. G, Packham M. A, Mustard J. F. Adenosine diphosphate-induced platelet aggregation in suspensions of washed platelets. Brit. J. Haemat 19: 7 1970;
  • 6 Emmons P. R, Mitchell J. R. A. Post-operative changes in platelet clumping activity. Lancet I: 71 1965;
  • 7 Bligh E. G, Deyer W. J. A rapid method of total lipid extraction and purification. Canad. J. Biochem 57: 911 1959;
  • 8 Bartlett G. R. Phosphorus assay in column chromatography. J. biol. Chem 234: 466 1959;
  • 9 Haslam R. J. Role of adenosine diphosphate in the aggregation of human blood platelets by thrombin and fatty acids. Nature (Lond) 202: 765 1964;
  • 10 Gronberg S, Caen J. P. Platelet aggregation in washed suspensions. Scand. J. Haemat 08: 161 1971;
  • 11 Packham M. A, Nishizawa E. E, Mustard J. F. Response of platelets to tissue injury. Biochem. Pharmacol. Suppl. 171 1968;
  • 12 Born G. V. R, Cross M. J. The aggregation of blood platelets. J. Physiol 168: 178 1963;
  • 13 O’Brien J. R. Changes in platelet membranes possibly associated with platelet stickiness. Nature (Lond) 212: 1057 1966;
  • 14 Rozenberg M. C, Holmsen H. Adenine nucleotide metabolism of blood platelets. IV. Platelet aggregation response to exogenous ATP and ADP. Biochim. biophys. Acta (Amst) 157: 280 1968;
  • 15 Packham M. A, Ardlie N. G, Mustard J. F. The effect of adenine compounds on platelet aggregation. Amer. J. Physiol 217: 1009 1969;
  • 16 Packham M. A, Evans G, Glynn M. F, Mustard J. F. The effect of plasma proteins on the interaction of platelets with glass surfaces. J. Lab. clin. Med 73: 686 1969;
  • 17 Nicholson J. T. Ph. D. dissertation. University of Lancaster; England: 1971
  • 18 Ireland D. M, Mills D. G. B. Detection and determination of adenosine diphosphate and related substances in plasma. Biochem. J 99: 283 1966;