It is well known that the function of platelets decreases progressively during storage of platelet concentrates at room temperature. To investigate this phenomenon in more detail, we have resuspended platelets that had been stored for 24 h or 72 h in fresh plasma, and we have measured the aggregation response and the ATP secretion. Conversely, the effect of plasma in which platelet concentrates (PC) had been stored for 24 h or 72 h, was tested on fresh platelets. Both the aggregation response to collagen and ADP and the collagen-induced ATP secretion of stored platelets partially recovered after incubation with fresh plasma (p <0.05). The same parameters measured with fresh platelets incubated in stored PC-plasma were found to be significantly reduced in comparison with the response of fresh platelets in fresh plasma (p <0.05). Finally, platelets were stored in a plasma-free medium, suitable for platelet storage and the supernatant was tested. This supernatant inhibited the function of fresh platelets in a storage time-dependent fashion. Boiling of these supernatants did not change the inhibiting capacities, whereas filtration over active charcoal did. Analysis of this supernatant revealed AMP and diadenosine tetraphosphate, which both inhibit platelet function.
These data show that stored platelets release nucleotides that inhibit platelet function in a reversible manner. This phenomenon may contribute to the decrease of platelet function during storage and the recovery of platelet function after transfusion.
REFERENCES
1
Di Minno G,
Silver MJ,
Murphy S.
Stored human platelets retain full aggregation potential in response to pairs of aggregating agents. Blood 1982; 59: 563-568
4
Shukla SD,
Morrison WJ,
Klatchko DM.
Response to platelet activating factor in human platelets stored and aged in plasma. Decrease in aggregation, phosphoinositide turnover, and receptor affinity. Transfusion 1989; 29: 528-533
8
Ishikawa Y,
Honda K,
Sasakawa S,
Hatada K,
Kobayashi H.
Prevention of leakage of di-(2-ethylhexyl)phthalate from blood bags by glow discharge treatment and its effect on aggregability of stored platelets. Vox Sang 1983; 45: 68-76
9
Watanabe E,
Tanaka K,
Sasakawa S.
Changes of platelet membrane associated Ca2+ during storage and by 37° C incubation with fresh plasma. Thromb Res. 1983; 32: 537-554
10
Sasakawa S,
Ishikawa Y,
Watanabe E.
Effects of storage and 37° C incubation with fresh plasma on platelet calcium distribution, mobilization, influx and aggregability. Thromb Res 1986; 42: 557-566
11
Ogawa A,
Ishikawa Y,
Sasakawa S.
Effects of changes in intracellular pH induced by platelet storage and treatment with monovalent cation ionophores on platelet functions. Thromb Res 1988; 52: 369-379
13
Fijnheer R,
Veldman HA,
van den Eertwegh AJM,
Gouwerok CWN,
Homburg CHE,
Boomgaard MN,
de Korte D,
Roos D.
In vitro evaluation of buffy coat derived platelet concentrates stored in a synthetic medium. Vox Sang 1991; 60: 16-22
14
Pietersz RNI,
Loos JA,
Reesink HW.
Platelet concentrates stored for 72 hours at 22° C prepared from buffycoats of CPD blood collected in a quadruple SAGM system. Vox Sang 1985; 49: 81-85
16
Coller BS,
Franza Jr BR,
Gralnick HR.
The pH dependance of quantitative ristocetin-induced platelet aggregation: theoretical and practical implications. A new device for maintenance of platelet-rich plasma pH. Blood 1976; 47: 841-854
17
Feinman RD,
Lubowsky J,
Charo I,
Zabinski MP.
The lumi-aggregometer: a new instrument for simultaneous measurement of secretion and aggregation by platelets. J Lab Clin Med 1977; 90: 125-129
18
de Korte D,
Haverkort WA,
van Gennip AH,
Roos D.
Nucleotide profiles of normal human blood cells determined by high-performance liquid chromatography. Anal Biochem 1985; 147: 197-209
19
de Korte D,
Haverkort WA,
Roos D,
van Gennip AH.
Anion-exchange high performance liquid chromatography method for the quantitation of nucleotides in human blood cells. Clin Chim Acta 1985; 172: 185-196
20
van Gennip AH.
Screening for inborn errors of purine and pyrimidine metabolism by bidimensional TLC and HPLC. In: Handbook of Chromatography, Nucleic Acids and Related Compounds, Vol. I Part A.
Krstulovic AM.
(ed)
CRC Press, Boca Raton, CA: 1987. pp 221-245
23
Harrison MJ,
Brossmer R,
Goody RS.
Inhibition of platelet aggregation and the release reaction by diadenosine polyphosphates. FEBS Lett 1975; 54: 57-60
27
Sakariassen KS,
Aarts PAMM,
de Groot PG,
Houdijk WPM,
Sixma JJ.
A perfusion chamber developed to investigate platelet inter-action in flowing blood with human vessel wall cells, their extra-cellular matrix, and purified components. J Lab Clin Med 1983; 102: 522-535
