To study the influence of contaminating leukocytes on the storage conditions of platelet concentrates (PC), various amounts of leukocytes were added to identical PC. From 12 blood donations, 12 leukocyte-poor PC were prepared and pooled. Subsequently, the pool was divided into 12 identical PC. The plasma volume of the PC was 58.6±0.6 ml, the platelet concentration was 1.01±0.04x10^9/ml (mean ± SD) and the red cell contamination did not exceed 10^7 per PC. To 4 groups of 3 PC, pooled leukocytes were added from the same 12 blood donations. The leukocyte contamination for each group of 3 PC was 0.14±0.05, 1.96±0.09, 5.53±0.98 and 13.0±0.93x10^6/ml (mean ± SD) for groups I-IV, respectively. The PC were stored for 7 days at 22 °C in normal polyvinylchloride bags. A significant correlation was found between increasing concentrations of leukocytes in the PC and the drop in pH (r=-0.93), glucose consumption (r=-0.91), lactic acid production (r=0.93) and release of lactate dehydrogenase (r=0.92) during storage of the PC. The excretion of β-thromboglobulin, depletion of platelet adenine nucleotides, decreased ability to incorporate 3H-adenosine into metabolic nucleotides and poor morphology of the platelets were also significantly correlated with an increased number of leukocytes in the PC. These data show that high concentrations of leukocytes in PC have a significant detrimental effect on the viability of platelets during storage at 22 °C. We conclude that for good storage conditions of PC, the upper limit of leukocytes per PC should not exceed 10^7.
Human blood platelets, prepared by the buffy-coat method, were prepared and stored in different synthetic media. In a synthetic medium based on gluconate, acetate and citrate (GAC), the pH was 6.8 on day 6. This medium was chosen for further evaluation. The total platelet count and the leukocyte contamination were significantly lower in the platelet concentrates (PCs) prepared in GAC compared to PCs prepared in plasma. Platelets stored in plasma or in GAC were equally functional when tested for aggregation and adenosine triphosphate (ATP) secretion. Only stimuli that act through the arachidonic-acid pathway induced a lower platelet response in GAC. Platelet morphology was quantified by measuring the difference in light transmission during stirring at different rates in an aggregometer; no significant differences for platelets stored in GAC as compared to plasma were observed. Activation of platelets was measured by binding of monoclonal antibodies (McAb) against the Gp Ilb/IIIa complex and against activation-dependent antigens (GMP140 from the a-granules and a 53-kD glycoprotein from the lysosomal granules). There was no difference in binding of these McAb between platelets prepared and stored in plasma or GAC. We conclude that platelets prepared by the buffy-coat method and stored in GAC have the same in vitro qualitities as platelets stored in plasma, except for the lower aggregation response by the archidonic-acid pathway. This is probably due to an acetate-induced decrease in intracellular pH.
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