These studies show that p-selectin expression on the platelet surface is a predictor of platelet viability, although the extent of shape change and the hypotonic shock response may be more sensitive.
Platelet concentrates stored for 5 days under currently optimal conditions at 22 C demonstrate approximately a 20-30% loss of viability as determined by CCI measurements in thrombocytopenic patients and radiolabeling studies with healthy volunteers. The functionality of platelets is substantially lost during storage. This loss appears, for most part, to be reversible upon infusion. In view of this, stored platelets should be acceptable for prophylactic treatment of thrombocytopenic patients in whom immediate hemostatic function is not critical. However, whether stored platelets are functionally adequate for immediate arrest of perioperative bleeding in surgical patients remain uncertain. Currently, a fully adequate and appropriate test that evaluates the hemostatic function of a stored platelet product is not available. In view of this, a desired goal in research and development of new platelet products may then be to attain a platelet product that is similar to fresh platelets in various haemostatic functions. Whether this is an obtainable or relevant goal remains to be seen.
Multiple laboratories were able to obtain comparable results with the ESC and HSR tests. They were able to show that the tests can be performed in an accurate, reproducible manner and with acceptable sensitivity.
The development of methods for storing platelet concentrates (PCs) at 22 degrees C for transfusion has been predominantly empiric, with minimal knowledge of metabolic events occurring during storage. It is known that a decrease in pH due to accelerated production of lactic acid in hypoxic conditions is a major cause for loss of platelet viability. In the current studies, we have measured metabolic parameters such as O2 and glucose consumption rates and CO2 and lactic acid production rates. We have also determined the O2 and CO2 transport capacities of various containers and the buffering capacity of plasma. The O2 consumption rate was 1.10 +/- 0.16 (SD) nmol/min/10(9) platelets. In well-oxygenated systems, lactic acid formation was 1.74 +/- 0.12 nmol/min/mL PC for PCs with a platelet count of 1 to 2 X 10(9)/mL; and 0.52 mol of glucose was consumed per 1 mol lactic acid produced. In a completely oxygen-free system, lactic acid production increased 5–8-fold. These calculations suggest that 85% of energy generation is derived through oxidative metabolism and that glucose may not be the primary substrate for this metabolism. Bicarbonate concentration, initially 22.1 +/- 1.6 mEq/L, decreased 1.41 +/- 0.18 nEq/min/mL PC for PCs with counts 1 to 2 X 10(9) platelets/mL. The loss of bicarbonate was caused by displacement by lactic acid and as a consequence of spontaneous CO2 efflux from the container. CO2 production, 2.3 +/- 0.4 nmol/min/10(9) platelets, was derived from oxygen consumption and the CO2 liberated from bicarbonate as it was consumed. A rapid fall in pH to levels below 7.0 (22 degrees C) took place when the bicarbonate concentration fell below 5 mEq/L as lactate concentrations reached 20 to 25 mmol/L. A further increase in lactate concentration from 25 mmol/L to 40 mmol/L correlated with a further fall in pH to 5.8. Thus, the ultimate storage life of a PC is determined by continuous lactate production and the fixed buffering capacity of plasma and by the glucose concentration of the PC. With knowledge of these parameters, methods for predicting pH as a function of time, platelet count, and O2 and CO2 transport capability of the container have been developed as guidelines for future work.
Regeneration of 2,3-diphosphoglycerate (DPG) was determined following transfusion of DPG-depleted group O red cells into group A recipients. Blood from five donors was stored in the adenine-containing solutions CPDA-1, AS-1 or AS-3 for 35 d at 4 degrees C. Post-transfusion red cell DPG and ATP were measured in separated group O red cells over a 7 d period. The studies confirmed rapid in vivo DPG regeneration with greater than or equal to 50% of the maximum level being achieved within 7 h. An average of 95% of the recipients' pre-transfusion DPG level was achieved by 72 h and by 7 d mean (+/- SEM) DPG levels relative to recipient's pre-transfusion DPG averaged 84% (+/- 13%), 92% (+/- 17%) and 84% (+/- 21%) for CPDA-1, AS-1 and AS-3 red cells, respectively. Results were comparable to those previously reported for blood stored in ACD for 15-20 d (Valeri & Hirsch, 1969; Beutler & Wood, 1969). The immediate regeneration rate, V, closely approximated first order regeneration kinetics with AS-3 red cells exhibiting double the rate of CPDA-1 red cells (P less than 0.001). AS-1 red cells exhibited an intermediate rate of regeneration which was not significantly different compared to either CPDA-1 or AS-3 (P greater than 0.05). V exhibited a significant (P less than 0.05) positive correlation with ATP levels 5-7 h post-infusion. ATP regeneration of the infused cells was rapid with a mean increase of 1.2 mumol/g Hb above post-storage levels being achieved 1 h following transfusion.
Irradiation has only a small effect on the properties of RBCs treated and stored according to the utilized protocols. Longer storage times after irradiation resulted in progressively reduced recovery while long-term survival remained unaffected.
An additive solution has been developed for storage of platelet concentrates (PC) which sustains improved in vivo and in vitro viability after 7 d of storage in second generation oxygen permeable containers. This platelet additive solution is a protein-free physiologic salt solution fortified with citrate, bicarbonate and glucose. The in vivo quality of the PC was evaluated by autologous radiolabelling with Indium-111-oxine to measure recovery and survival by multiple hit analysis. The in vitro quality was evaluated by total ATP content, hypotonic shock response and extent of shape change with ADP. Ten paired studies were performed with PC from the same donor being stored for 7 d at 22 degrees C in both CPDA-1 plasma and the additive solution. Mean recoveries and survivals were found to be substantially higher with PC stored in the additive solution than with PC stored in CPDA-1 plasma (51.0 +/- 7.8% and 144.1 +/- 15.9 h versus 36.6 +/- 10.7% and 110.4 +/- 31.6 h). The differences were statistically significant (P less than 0.001). The results of the in vitro assays described above parallelled the in vivo results, with statistically significantly superior results (P less than 0.01) for all parameters of PC stored in the additive solution. This study is the first to show that PC quality may be improved and storage extended using an additive solution.
A new, in-line high-efficiency 3-5 log10 leucodepletion filter system (Leukotrap RC system) was used to investigate the effect of pre-storage white cell removal on the quality of AS-3 red cell concentrates stored for 42 d at 4 degrees C. Median residual white cell content was 4 x 10(5) when filtration was performed at 22 degrees C within 8 h of phlebotomy (n = 20) and 3.2 x 10(4) when filtration was performed at 4 degrees C 12-24 h after phlebotomy (n = 24). None exceeded 1 x 10(6) WBC per red cell product. Filtration was rapid (median 28 min), and red cell loss averaged (mean +/- 1 SD) 6.4 +/- 0.7%. In a paired study design, post-transfusion recoveries of 42 d stored red cells in the filtered units averaged 84 +/- 6% v 82 +/- 8% for unfiltered units (P < 0.05) and post-storage haemolysis. ATP, osmotic fragility, K+ and pH were significantly (P < 0.05) better in the filtered units. Reduced glycolytic activity was also observed in the filtered units, and there was a correlation between osmotic fragility, glucose consumption, and lactate produced in standard units that was not present in leucodepleted units. In conclusion, this study suggests that leucodepletion of AS-3 red cell concentrates prior to storage results in better maintenance of the integrity of the red cell membrane with reduced glycolytic activity. There was a modest improvement in post-infusion viability sufficient to offset the filtration-induced loss and to result in an equivalent red cell product.
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