Analysis of Platelet Factor 3 in Platelet Concentrates Stored for Transfusion

The amount of platelet factor 3 (PF3) activity expressed in stored platelet concentrates (PC) was measured in conjunction with extracellular LDH levels. Standard manual techniques for preparation of PC resulted in PF3 and LDH levels remarkably higher than those observed in PC prepared by apheresis or special manual plateletpheresis. During storage of PC, PF3 activity rose 2- to 10-fold, while LDH levels rose less than 2-fold over starting values. Loss of LDH and appearance of PF3 expressed as a percent of total per platelet were significantly correlated only in standard, manual PC. Approximately half of the PF3 activity observed in any type of PC remained in the supernatant plasma after centrifugation. Upon gel filtration, the supernatant PF3 activity eluted in a high molecular weight peak containing phosphate and light-scattering material. Our findings indicate that platelets in standard, citrated PC express PF3 in amounts that approach that of frozen-thawed (lysed) platelets; however, the manner in which the PF3 activity appears suggests that stored platelets undergo a combination of activation and damage processes.     

Metabolic Status of Platelet Concentrates during Extended Storage: Improvement with Pharmacological Inhibitors and Reduced Surface-to-Volume Ratio

The depletion of plasma nutrients and buffering capacity may present a potential barrier to the long-term liquid storage of platelet concentrates (PC). We have found that PC prepared with reversible inhibitors of platelet activation added to the citrate anticoagulant and stored at a reduced surface-to-volume (S/V) ratio have a much slower rate of lactate build-up (p less than 0.01), slower consumption of glucose (p = 0.05), and more stable pH (p less than 0.01) than controls. By pO2 and pCO2 measurements, PC prepared with inhibitors showed evidence of continued respiration and responsiveness even after storage at 22 degrees C for 15 days. In addition, these PC released only 11% of the total cellular LDH during the storage period as compared to the release of 43-67% of the total LDH in control PC. Maximum benefit of the inhibitors was seen after reduction of the S/V ratio of the storage container, which was made possible by the reduced metabolic demands of platelets stored in the unactivated state. These data suggest that the fall in pH and loss of platelet integrity associated with the platelet storage lesion are correlated with a high metabolic rate which can be controlled by inhibiting the activation of platelets during preparation and storage. The use of these inhibitors and reduced bag surface area may make prolonged liquid storage of platelets feasible.     

Platelet storage at 22 degrees C: effect of type of agitation on morphology, viability, and function in vitro.

Recovery in vivo after 51Cr labeling, platelet morphology, and platelet aggregation were studied with platelet concentrates (PC) stored for transfusion under carefully controlled conditions. PC were prepared to a final volume of 50 ml from whole blood anticoagulated with citrate-phosphate-dextrose (CPD). The platelet count was kept between 0.8 and 1.6 X 10(12) platelets/liter. The PC were stored in bags constructed of polyvinylchloride (PVC) or polyethylene (PE) at 22 degrees C for 72 hr. The bags were placed on a horizontal shaker or a ferris wheel for agitation during storage. No significant changes in pH or platelet count were observed during storage. PC stored on the wheel showed moderate loss of viability and a marked deterioration of platelet morphology and aggregation compared to the shaker. PC stored on the shaker in bags made of PE showed better aggregation with ADP and thrombin but had the same viability and morphology as PC in bags constructed of PVC. Maintenance of normal platelet morphology as determined by phase-contrast microscopy, extent of shape change response, and the size distribution according to the Coulter Counter correlated with recovery in vivo.     

Effect of Filtration on Subsequently Stored Platelet Concentrates

The effect of filtration on the quality of platelet concentrates (PC) during storage was investigated. Two leukocyte depletion filters (Pall PL50HF and Sepacell PL-10A) were applied to filter PC made from a pool of 4 buffy coats. For each experiment 3 PC were pooled and divided into 3 identical PC to eliminate differences between the PC. Two PC were filtered, and the third PC served as an unfiltered control. A total of 12 experiments was performed. Before filtration, volumes of the PC were 263±11.7 ml (mean±SD). Platelet and leukocyte counts per PC were 241±25.9 times 10⁹ and 7.2±1.8 times 10⁶, respectively. After filtration leukocyte counts did not exceed 5 times 10⁴ in any of the PC. In the PC filtered with the Pall PL50HF the mean platelet loss was approximately 14% and with the Sepacell PL-10A, 17%. During a 9-day storage period the pH, PO₂, PCO₂, bicarbonate, lactate and glucose concentration and LDH release as well as the morphology, examined by the swirling effect and microscopically, were not significantly different in filtered and unfiltered units. Filtration through the 2 investigated leukocyte depletion filters for PC did not adversely affect in vitro viability of the platelets during storage.     

Platelet storage at 22 degrees C: role of gas transport across plastic containers in maintenance of viability.

Containers constructed of polyvinylchloride (PVC) are used for the storage of platelet concentrates (PC) for transfusion, At 22 degrees C, pH often falls to such low levels (pH is less that 6.0) that viability is lost. Far lesser degrees of pH fall are observed in bags constructed of polyethylene (PE). In this study, pH, PO2, PCO2, platelet count, lactate concentration, microscopic morphology, and viability after 51-chromium labeling were evaluated during storage at 22 degrees C under a variety of circumstances. The results indicate that (1) pH falls because of the generation of lactic acid by platelet glycolysis and, under some circumstances, the retention of CO2. (2) Rate of pH fall is, therefore, roughly proportional to the platelet count. (3) PE is more permeable to gases, thereby allowing CO2 escape from and easier O2 entry into the stored PC; the higher O2 tensions suppress glycolysis by the Pasteur effect. (4) Adequate agitation and container size are critical if the beneficial effect of PE is to be obtained. (5) In general, platelets stored in PE containers have excellent viability in vivo although CO2 escape can result in elevations in pH which are deleterious. (6) Storage in a 10% CO2 atmosphere prevents these deletrrious pH elevations without otherwise impairing platelet viability; (7) Results similar to those achieved with PE can be achieved with PVC if this material is made thinner to allow easier penetration of gases     

Platelet metabolism during storage of platelet concentrates at 22 degrees C

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.     

Coagulation Factor Activation in Stored Platelet Concentrates is Modulated by the Platelets

To assess alterations in coagulation proteins in stored platelet concentrates, we used a series of platelet parameters and measures of coagulation activation to compare samples collected before unit donation, during the processing of platelet concentrates (PC) from CPDA-1 blood, and in storage up to 5 days as well as stored platelet-poor plasma (PPP). Storage-dependent increases in activated partial thromboplastin time and prothrombin time were seen in both PC and PPP. However, FVII, FXI, FXII, kallikrein activity and prothrombin F1.2 levels remained unchanged in stored PC. Surprisingly, in stored PPP, significant alterations in FXII, FVII, kallikrein and prothrombin F1.2 levels were seen. Platelet morphology and surface marker studies demonstrated platelet activation during storage. These data suggest that the presence of platelets in the CLX storage container partially suppresses coagulation activation at significant cost to platelet viability.     

Platelet size analysis in the quality assurance of platelet concentrates for transfusion

Summary The platelet distribution width (PDW) as analysed on standard haematology cell counters is an indicator of size dispersion in the platelet population. Using a Sysmex® E-2500 analyser platelet concentrates prepared for transfusion showed an increase in PDW over storage. This increase correlated strongly with in vitro indicators of platelet viability (pH and response to osmotic stress). PDW may thus be useful for clinical haematology laboratories as a predictor of the viability of transfused platelets. The same instrument gave a measure of the largest platelets in the platelet population as a large cell ratio (P-LCR). For platelet concentrates with less than 8 × 10¹⁰ platelets/unit, the P-LCR at preparation was negatively associated with the end of storage pH, indicating that the presence of large platelets increases the production of lactic acid and accelerates the platelets’ metabolic storage lesion. This information may be useful in determining storage conditions for single donor platelets harvested by apheresis.     

Platelet Storage Lesion in Second-Generation Containers: Correlation with Platelet ATP Levels

The nature of platelet lesion occurring with storage of platelet concentrates (PC) in second-generation containers was investigated using various storage media and storage periods up to 14 days. In CPD-plasma (control medium), the changes which occurred progressively during storage were loss of discoid shape, microscopic platelet aggregate formation, fragmentation and the appearance of disintegrated, 'balloon' forms. By day 14 less than 10% of the platelets were discoid in shape, the platelet count had decreased by 23%, and there was a 5-fold increase in the amount of lactate dehydrogenase in plasma. Associated with this was a decrease in the platelet oxygen consumption rate, D(O2), loss of cellular ATP and extent of ADP-induced shape change, and a decrease in the hypotonic shock response. These parameters decreased at a similar rate, with a 50% decrease (t1/2) at days 7-9. They correlated highly with each other during storage and also with a fall in pH. At day 14 of storage, mean pH was 6.1 +/- 0.3. To evaluate the effect of pH stabilization during storage, 4 mEq sodium bicarbonate was added to PC in CPD-plasma. Although pH maintenance was much improved, 7.2-6.6 during 14 days of storage, the same in vitro lesions developed, although more slowly. The t1/2 of the same parameters was prolonged for approximately two days. When PC were stored in a plasma-free physiologic salt solution whose salt composition was similar to CPD-plasma, the t1/2 of the parameters increased to 11-15 days of storage, although the platelets eventually developed the same in vitro lesions.(ABSTRACT TRUNCATED AT 250 WORDS)     

Platelet fragments do not contribute to elevated levels of platelet associated IgG

Most assays that measure platelet associated IgG (PAIgG) relate the IgG associated with the test platelets to the platelet count. This could lead to a systematic error if platelet fragments were present in the washed platelet sample but not counted. To address this issue, we studied platelets from patients with idiopathic thrombocytopenic purpura (ITP), thrombocytopenia complicating cardiopulmonary bypass, and laboratory synthesized platelet fragments (freeze-thawed) using electron microscopy. Scanning electron microscopic (SEM) examination of the platelet samples demonstrated appreciable numbers of fragments only in the freeze-thawed specimens. Yet, 'fragments' could be seen in all specimens using transmission electron microscopy (TEM). Most of these 'fragments' proved to be artefacts: we found that the ratio of 'fragments' to intact platelets observed in the TEM specimens was similar to the estimated ratio of 'fragments' to platelets that would have been generated had the specimens been sectioned at 90 degrees to the plane of the actual section. Platelets labelled with the membrane label 125I-iodosulfanilic acid were fragmented by repetitive freeze-thawing. Measurement of pellet radioactivity following washing indicated that the fragments were lost during the washing procedure. These studies indicate that elevated levels of platelet associated IgG in ITP do not represent artefacts due to contamination of the test platelets by platelet fragments.     

Platelet Storage Lesions in Second-Generation Containers: Correlation with in vivo Behavior with Storage up to 14 Days

The relationship between in vivo behavior and in vitro characteristics of 59 platelet concentrates (PC) stored for up to 14 days in a synthetic medium or in CPDA-1 plasma was systematically investigated. 25 paired studies (1 study was incomplete) were performed comparing platelets suspended either in the synthetic medium or CPDA-1 plasma with 5 days (n = 5); 7 days (n = 10); 10 days (n = 5); and 14 days (n = 5) of storage. In addition, 10 control studies were performed with freshly prepared PC (6-24 h) in CPDA-1 plasma. Both percent recovery and survival estimations showed decreases with increasing storage duration, irrespective of storage medium used. In both media, with prolonged storage, the platelet survival curves not only became shorter, but also increasingly exponential, suggesting that in vitro storage caused progressive damage to the platelets present in circulation. Survival curves of platelets suspended in synthetic medium remained more linear, indicative of less random damage during storage. Mean population lifespan (MPL) of the stored PC was determined by the area below the survival curve divided by the mean percent recovery for the fresh PC, which was 55%. MPL decreased from 4.5 days (fresh PC) to 0.4 days after 14 days of storage in plasma, with a 50% reduction (t1/2) estimated at 7.2 days of storage. MPL t1/2 for PC stored in the synthetic medium was estimated to be 8.8 days.(ABSTRACT TRUNCATED AT 250 WORDS)     

Storage of platelet concentrates using ion exchange resin charged with dibasic phosphate.

Platelet concentrates were prepared at twice the normal concentration and stored at room temperature for 7 days in either standard bags (controls) or bags to which 1 or 2 g of Amberlite resin beads charged with dibasic phosphate had been added. The resin beads served as a buffer system by providing a "slow release" form of phosphate ions as well as by binding CO2 produced during platelet metabolism. Control platelets demonstrated rapid falls in pH, ATP content, morphology score, and thrombin-induced nucleotide release after 24 hr of storage with a fall in pH to less than 6.0 by day 3. Profound ultrastructural changes and a rise in pO2, suggesting loss of platelet viability, accompanied these changes. In contrast, the resin-stored platelets remained near normal after 24 hr of storage, with preservation of discoid morphology, 95% of ATP levels, excellent ultrastructural appearance, and evidence of continued oxygen consumption after 3 days of storage. Even after 7 days of storage, ATP levels remained greater than 50% of baseline and ultrastructurally intact platelets were seen. In the 1-g resin bags the pH remained at baseline levels (6.9-7.0), while there was a rise in pH in the 2-g resin bags. These results demonstrate the beneficial effects of maintaining a higher pH during platelet storage and provide a new approach to studying the metabolic changes that occur during longer term storage.     

The effect of acidified glucose nutrient solution on preservation of platelet concentrate

Summary The preservation effect of a new platelet solution-acidified glucose nutrient solution (AGN)-was tested by adding AGN to pooled platelet rich plasma (PRP) prior to preparation of platelet concentrates (PC). Each PRP unit was prepared from a unit of whole blood and four PRP of the same blood group were pooled in 400 ml volume PVC bags. Equal aliquots of each pooled PRP were made prior to preparation of PC. AGN was added to one aliquot and nothing was added to the control aliquot. Equal volume and concentration PC were then prepared and the PC were further aliquoted for storage at 22°C. After five days, the parameters of platelet count, pH, aggregability and hypotonic shock response (HSR) for PC preserved in AGN plasma were better than those of the controls preserved in normal ACD plasma. The aggregability and HSR of AGN platelet concentrates recovered to baseline after two h of incubation with fresh plasma. The results of electron microscopy show that platelets preserved in AGN have less changes in morphology. The results of our work suggest that the preservation effect of AGN on PC is similar to the effect of using second generation containers or preparation of platelets from thrombocytapheresis.     

Platelets Stored in a Glucose-Free Additive Solution or in Autologous Plasma – An Ultrastructural and Morphometric Evaluation

We evaluated a new glucose-free citrate-acetate-NaCl platelet additive solution (PAS 2). In series I, platelet concentrates (PC) were prepared by apheresis and subsequently stored either in plasma (n =16) or in PAS 2 (n =15). In series II, PCs were prepared from pools of four buffy coats (BC) and stored in plasma (n =12) or in PAS 2 (n =11). By means of ultrastructural morphometry, the volume fractions of α-granules, the open canalicular system (OCS) and the fraction of storage granules secreted into the OCS were analyzed during storage for up to 8 days. Additionally, we determined pH, glucose, lactate, pCO₂, HCO₃^–, lactate dehydrogenase and platelet factor 4. Apheresis platelets stored in plasma showed no changes in their contents of α-granules and in the fractions of the OCS. In contrast, apheresis platelets stored in PAS 2 displayed a decrease of their relative volume fraction of α-granules from 9.1±1% on day 1 to 3.7±0.9% on day 5. The fraction of the OCS increased from 7.4±0.8% on day 1 to 17.1±1.4% on day 3. On day 8, 93±9% of all platelets were lysed. Levels of glucose were significantly lower in these preparations and after day 3 glucose consumption decreased to zero. Among PC derived from pooled BC, differences between storage in PAS 2 or plasma were less striking. Only the fraction of α-granules secreted into the OCS was significantly greater in BC derived PC stored in PAS 2 on all days. These PCs stored in PAS 2 had a higher plasma carryover (30%) in comparison to apheresis PC stored in PAS 2 (10%). We conclude that plasma is superior to PAS 2 for storage of both apheresis and buffy coat platelets. For preservation of the structural integrity of platelets, the use of PAS 2 requires a minimum of 30% plasma carryover.     

Evaluation of refrigerated platelet concentrates supplemented with low doses of second messenger effectors

With the goal of producing haemostatically effective platelet concentrates (PCs) with a longer shelf-life, we aimed to identify a simple combination of platelet inhibitors, with a low pharmacological load, which could avoid the unacceptable loss of platelets stored under refrigerated conditions. PCs stored with different combinations of second messenger effectors were analysed at days 5, 10 and 15 of storage and compared with those supplemented with ThromboSol--a combination of six platelet inhibitors that protects cells from cold damage. The following parameters were analysed: platelet counts, biochemical parameters (glucose, pH, bicarbonate, lactate), cell lysis (lactic dehydrogenase, LDH), membrane glycoproteins (GPs), platelet aggregation, fibrinogen binding and hypotonic shock response. We characterized the combination of amiloride and sodium nitroprusside (at 1/2 the dose included in ThromboSol). This was found to be similar to ThromboSol and superior to nontreated units in the prevention of cold-induced platelet aggregation at day 15 of storage (maintenance of 78% and 80% of initial platelet counts, respectively), preservation of GPIbalpha (11% and 12% better maintenance of mean fluorescence intensity compared with control units, respectively), and reduced cell lysis (13% and 11% decrease in supernatant LDH, respectively). The reduced pharmacological load with the identified solution compared with ThromboSol is an argument in favour of the potential use of these agents when designing strategies to improve PC storage.     

Irradiated Blood Platelet Concentrates Stored for Five Days -Evaluation by in vitro Tests

Abstract. Platelet concentrates, irradiated with 15 Gy and stored for 5 days at room temperature under standardized conditions, were evaluated by in vitro tests and electron microscopy, in a paired study with nonirradiated platelets from the same concentrates, to investigate their usefulness for transfusion. The results showed no significant differences between the two groups in pH, pO₂, PCO₂, in vitro platelet aggregation, LDH, β-thromboglobulin and thromboxane-B₂. Examination by electron microscopy showed a higher degree of degranulation in the 5-day-old platelets but no certain difference between irradiated versus nonirradiated platelets. On the basis of satisfactory in vitro storage properties, platelet concentrates can be stored for 5 days in PL-1240 bags after irradiation. However, we recommend irradiation just before transfusion whenever possible.     

Platelet Collection with the Autopheresis-C® Apheresis System

This report describes a new system for collection of platelet concentrate (PC) and cell-free plasma (PPP) from apheresis donors. The system uses two separation devices and requires only a single venipuncture. The Plateletcell device separates primary platelet concentrate (PPC) from anticoagulated whole blood and the Plasmacell-C device separates the PPC into PC and PPP. Results of functional studies performed indicate that the separation process does not alter viability of either the PPC, the PC, or the PPP. Platelet function after 5 days of storage is maintained. An average yield of 3.4 +/- 0.7 x 10(11) platelets in 201 g of PC and 422 g of PPP were harvested in 71 +/- 13 min of donor time from donors with preprocedure hematocrits averaging 42.5 +/- 2.0% and preprocedure platelet counts averaging 265 +/- 61 x 10(3)/microliters.     

Role of Lactate in Platelet Storage Lesion

It is known that lactate accumulation may cause a pH fall in platelet concentrates (PC) during storage, and this phenomenon causes platelet morphological lesions and loss of platelet in vivo viability. In this study, we added increasing amounts of lactate to identical PC in order to evaluate the role of hydrogen ion accumulation in determining platelet activation and lesion during storage. Six hours after PC preparation, lactate was added to PC1 and PC2 at 20 and 12 mM final concentrations, respectively, while PC3 served as control. In PC1, pH was lower than 6.3, and platelet function and discoid morphology were lost. PC2 were stored for 7 days at pH values ranging from 6.4 to 6.6, and most results of in vitro measurements reflecting platelet function such as osmotic reversal, ATP release and aggregation in response to different stimuli were not significantly inferior when compared to controls. The addition of lactate had no apparent effect on the rise of platelet activation markers P-Selectin, lysosome-like protein gp 53, platelet-bound fibrinogen and granulophysin, while a reduction of borderline significance was observed in glycoprotein Ib expression after pH reduction to values lower than 6.6. It is concluded that the rise of platelet activation markers during storage reflects platelet lesions different from those determined by lactate per se.     

Platelets Stored in a New-Generation Container

Background and Objectives : In order to preserve platelet concentrates (PCs) with high yields, a new polyolefin platelet storage container (PL 2410, 1.3L, Baxter, La Châtre, France) with increased gas permeability in combination with a larger surface area has been developed. The storage capacity was studied with platelets in plasma and platelets additive solution. Materials and Methods : Platelet concentrate pools (PCs) of different yields suspended in either plasma (PCs-PL; n = 30) or PAS II (PCs-PAS; n = 37) were prepared. For preparation of PCs with a low, intermediate and high number of platelets 3, 5 and 6 buffy coat (BCs) were pooled with different volumes of plasma and 5 and 6 BCs were pooled with different volumes of PAS II, in order to obtain PCs of equal volumes comparable with routine conditions. All PCs were stored on a flatbed shaker at 22±2°C and evaluated on days 1, 2, 5 and 7 by determining platelet and white cell counts, pH (37°C), pO₂, pCO₂ and swirling score. Results : Platelet yields ranged from 1.5 up to 5.5 × 10¹¹ platelets/U. On day 7 all PCs-PL (n = 4) with platelet yields above 4.5×10¹¹ had a pH value below 6.8 (range 5.91–6.79). While 7 of 8 PCs-PAS units with platelet yields above 4.0×10¹¹/U showed a pH value below 6.8 (range 6.31–6.70). Conclusion : During 7 days of storage in a new 1.3-liter platelet container, the pH was maintained above 6.8 in PCs in plasma with a yield between 1.5-and 4.5×10¹¹/U; when PAS II was used, the maximum platelet yield allowed for comparable pH maintenance was somewhat lower (4.0x10¹¹/U).     

Platelet concentrates irradiated with ultraviolet light retain satisfactory in vitro storage characteristics and in vivo survival

We have previously reported that platelet concentrates (PC) may be irradiated with ultraviolet light (UVL) in a cryostorage pack such that mixed lymphocyte reactions (MLR) are abolished whilst satisfactory platelet function is retained during subsequent storage using Fenwal PL-1240 containers. We have now studied both platelet structure and function after irradiation in DuPont Stericell bags which are both UV-permeable and biocompatible. The irradiation dosage was 3000 Joules/m2 of UVL at a mean wavelength of 310 nm; a dose previously shown to abolish MLR. No detriment to platelet function was observed when compared to control as measured by aggregation responses to adenosine diphosphate (ADP), collagen and ristocetin, hypotonic shock response and pH during 5 d of storage. Lactate levels were significantly higher (P less than 0.01) and glucose levels lower (P less than 0.01) in UV-treated PC, although in the majority the lactate level did not exceed 20 mmol/l. Betathromboglobulin and platelet factor 4 levels were higher during storage in the UV group, the latter reaching significance (P less than 0.05). When whole platelets and platelet membranes were stained with Coomassie blue or Periodic Acid-Schiff's reagent after electrophoresis in polyacrylamide gels no obvious alterations to major membrane constituents were observed on days 1 and 5 of storage. Paired in vivo autologous studies in healthy volunteers using 111Indium-labelling were performed at the end of 5 d of storage. The UV-treated platelets gave satisfactory and equivalent results for recovery, half life and survival when compared to controls. We conclude that PC irradiated with UVL and stored for 5 d in DuPont Stericell containers appear suitable for transfusion and may prove to be nonimmunogenic. Further in vivo studies of haemostatic efficacy and recipient alloimmunization are now warranted.