Detecting failed WBC-reduction processes: computer simulations of intermittent and continuous process failure.

BACKGROUND: By regulation, ongoing process control of WBC-reduced processes is performed on 1 percent of WBC-reduced components, typically four to five samples per month. However, prospective study of the power of this small sample has been difficult. Using computer-generated "residual WBC" distributions, sample size sensitivity to continuous or intermittent WBC-reduction failure was examined. STUDY DESIGN AND METHODS: Populations of log-normally distributed values (mean +/- SD, 4.5+/-0.5; n = 10(5)) were generated. Continuous failure (log-normality maintained) was simulated by incrementally increasing the population mean or its SD. Intermittent failure (bimodal distributions with discrete subpopulations of WBCs > the FDA cutoff) was simulated by admixing increasing percentages of secondary outlier populations. Sample sizes of 4 to 60 were examined (500 repetitions each) for their power to detect drift or failure by standard control criteria. RESULTS: Normally distributed low variance failure was easily detected by comparison of the mean of four samples to an upper control limit (95% confidence of detecting 2% failure). However, 40 samples were required to detect > 5 percent intermittent (bimodal) failure or high variance failure with 90-percent confidence, and only if individual WBC values were compared to cutoff. CONCLUSION: Sampling error limits the detection of high variance or bimodal distributions. While the mean of a small sample is highly sensitive to shifts in a low-variance normal distribution, the detection of a high-variance bimodal population requires a large number of individual values compared to cutoff. Therefore, the number of samples required for confident failure detection depends on both the nature of the underlying distribution and the interpretive criteria. Further research is necessary to determine the true distributions of WBC-reduction process failure, as well as clinically relevant quality limits.

Detecting failed WBC-reduction processes: a quality assurance program introduced in a blood center.

BACKGROUND: Pragmatic yet statistically valid quality assurance (QA) programs are necessary so that blood centers can select, validate, and monitor their WBC-reduction processes. A QA system for WBC-reduction processes based on the practical application of statistical theory within a large blood center was developed. The system identifies parameters for procedure and component evaluation and provides sample size and formatting suggestions. STUDY DESIGN AND METHODS: Analyses of both procedure and component performance were undertaken during the purchase, validation, and control of filtration and apheresis WBC-reduction processes at Blood Centers of the Pacific from 1997 through 1999. QA analysis was categorized on the basis of whether the process was new to the organization or was a modification of a previously validated system. The numbers of samples necessary to consistently detect failure in platelet yield, unit volume, pH, and WBC count was statistically determined by parametric and nonparametric techniques. RESULTS: Parametric analysis (power analysis) of the mean +/- SD of smaller numbers of samples was highly sensitive to shifted distributions, but only if the shift was normally distributed. Nonparametric analysis, necessary when the nature of the underlying distribution is unknown, suggested a minimal sample of 40 was required to achieve high confidence that significant bimodal failure (a secondary population with WBCs 5% above the cutoff) would be detected. CONCLUSION: A QA system, developed for the evaluation of new or revised WBC-reduction processes, was based on statistical analysis of normally and non-normally distributed process failure. The number of samples was determined that allowed the achievement of confidence and tolerance levels considered appropriate within the blood center. Suggestions for outlier evaluation and a format for performance documentation have also been developed. To better define blood center quality goals, further research is necessary on donor and component biologic variability and the most significant modes of WBC-reduction process failure.

Viability and in vitro properties of AS-1 red cells after gamma irradiation.

BACKGROUND: Irradiation has been shown to adversely affect both in vivo 24-hour recovery (recovery [%]) and in vitro properties of stored red cells (RBCs). There is uncertainty as to how these changes are related to the day of irradiation and the length of storage after irradiation. STUDY DESIGN AND METHODS: Four protocols used day of irradiation and storage time after irradiation as the independent variables. At the conclusion of the storage period, viability was measured with radiolabeled RBCs as the recovery and the long-term survival time for RBCs that were circulating beyond 24 hours. In addition, in vitro values including RBC ATP, hemolysis level, and supernatant potassium were measured. Each subject donated 2 units of whole blood (CPD) and received autologous irradiated and untreated control RBCs (AS-1) on two separate occasions. RESULTS: Reduced recovery in irradiated units was noted when compared to that in control units, and the reduction was most apparent with long periods of storage after irradiation, irrespective of the day of irradiation. With irradiation on Day 1 of storage and a total storage period of 28 days, mean +/- SD recovery (single label) was 84.2 +/- 5.1 percent for control RBCs and 78.6 +/- 5.9 percent for irradiated RBCs (n = 16; p<0.01). With irradiation on Day 14 and storage through Day 42, the recoveries were 76.3 +/- 7.0 percent for control RBCs and 69.5 +/- 8.6 percent for irradiated RBCs (n = 16; p<0.01). Less reduction in recovery was observed with shortening of the postirradiation storage time. When the total storage period was reduced to 28 days after Day 14 irradiation, the recoveries were not significantly different. With an additional 2-day storage period after irradiation on Day 26, the recoveries were also comparable. Long-term survival times for control and irradiated RBCs were not significantly different in any of the four protocols. RBC ATP levels and hemolysis were minimally, but significantly influenced by irradiation. Supernatant potassium levels, however, were substantially increased after irradiation in each of the four protocols. CONCLUSION: 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.

Clinical trial and local process evaluation of an apheresis system for preparation of white cell-reduced platelet components.

BACKGROUND: A new method for the consistent preparation of white cell (WBC)-reduced plateletpheresis components, the Spectra Leukoreduction System (LRS), was evaluated by clinical trial and local process validation. The centrifuge-based system was projected to decrease the WBC content of plateletpheresis components to a level below 1 x 10(6) per unit. Phase I and II clinical trials were performed. The manufacturer's claims were then tested at the local level with an ongoing quality assurance program. STUDY DESIGN AND METHODS: In Phase I, a cross-over analysis of five subjects compared LRS to standard plateletpheresis procedures in collection efficiency and component quality: a panel of in vitro measures was taken on Day 0 and Day 5. In Phase II, the LRS process was tested on a larger scale (n = 57; control = 58) with component transfusion. Finally, validation, determination of degree of conformance with standards, and ongoing quality control were performed locally on a newly installed instrument. RESULTS: Phase I and II trials revealed no significant differences between LRS and control units in donor or recipient safety and comfort, platelet function and yield, or component volume. WBC per-unit values were significantly different: the LRS median per unit was 3.2 x 10(4) WBCs, versus 81.4 x 10(4) for control units. Assessment of process capability gave an estimate of 99-percent confidence that 99.5 percent of LRS units would be WBC reduced to < 1 x 10(6) WBCs. Local process validation and quality control revealed 90-percent confidence that 99 percent of the units would be WBC reduced and 99.9-percent confidence that 75 percent would exceed platelet yield standards; the process was stable over time. CONCLUSIONS: The LRS is safe for apheresis and the component produced is safe for transfusion with platelet function and yield equivalent to controls and WBC reduction superior to controls. Local process evaluation confirmed that component quality meets the goals of the institution.

Automatic volumetric capillary cytometry for counting white cells in white cell-reduced plateletpheresis components.

BACKGROUND: As the benefits of white cell (WBC)-reduced blood components become increasingly apparent, the need has arisen for a simple, automated WBC-counting technique that is sensitive to low WBC concentrations. Automated volumetric capillary cytometry was evaluated for its ability to quantify residual WBCs in WBC-reduced plateletpheresis components. STUDY DESIGN AND METHODS: The volumetric capillary cytometry system evaluated uses a laser to excite fluorescent dye-labeled nucleated cells. The number of nucleated cells per microliter is reported. Four studies were performed: linearity, precision of results near the value of 5 x 10(6) WBCs per unit, the limit of detection, and correlation to the Nageotte manual counting method. RESULTS: Assay values correlated to expected values (range, 0-125 WBC/microliter) with an r2 > 0.99. In the range of 5 x 10(5) WBCs per unit the CV was 8.5 percent, and concentration differences of 0.15 log10 were detectable. The limit of detection was 1.0 WBCs per microliter (95% upper confidence limit). The assay correlated to the Nageotte method with an r2 of 0.98, slope of 1.0, and y-intercept of 2.0 WBCs per microliter. Assay results were 10 to 15 percent higher than Nageotte results, in samples with values near 5 x 10(6) WBCs per unit. Technician time per sample was 2 to 3 minutes. CONCLUSION: Volumetric capillary cytometry is precise and sensitive to small differences in WBC concentration in the range of clinical interest. The device provides an efficient new method for quality assurance and control of WBC-reduced plateletpheresis products.

In vitro and in vivo effects of prestorage filtration of apheresis platelets.

BACKGROUND: The effect of prestorage filtration on the quality of apheresis platelet concentrates stored for transfusion is undetermined. STUDY DESIGN AND METHODS: Investigation of 11 plateletpheresis components used a concurrent paired-study design. On the day of collection, each component was equally divided into two suspensions; one half was filtered, and the other half was not. Each suspension was stored for 5 days. In vitro testing was performed on the day of collection (Day 0) for cell counts and on Day 5 for measurements of lactate, glucose, blood gases, pH, platelet ATP, hypotonic stress ratio, extent of shape change in response to ADP, tissue necrosis factor alpha, interleukin 8, interleukin 1 alpha, interleukin 1 beta, interleukin 6, and platelet surface glycoproteins by flow cytometry. At the end of the 5-day period, a sample was taken from each of the two suspensions, radiolabeled with either 51Cr or 111In, and transfused concurrently. Posttransfusion samples were drawn for measurements of recovery and platelet survival and for functional assessment of the ex vivo ability of the circulating radiolabeled platelets to aggregate in response to ADP. RESULTS: The apheresis component had a mean platelet yield of 3.2 +/- 0.4 x 10(11) and a white cell yield ranging from 1 x 10(5) to 1 x 10(8), with a median of 2 x 10(7). Filtration resulted in a platelet loss of approximately 10 percent and a variable 2 to 3 log10 reduction in white cell content. No significant differences between filtered and unfiltered suspensions in paired t tests that would likely have an impact on platelet quality were observed in the in vitro tests. The in vivo recovery and survival were highly similar and not statistically different in filtered and unfiltered paired suspensions: the mean difference was 1.2 +/- 4.0 percent for recovery and 7.0 +/- 15 hours for survival. The functional assessment by aggregation to ADP showed no difference between filtered and unfiltered suspensions. A small decrease in tumor necrosis factor alpha and interleukin 8 was evident in the filtered suspension as compared to levels in the unfiltered suspensions. CONCLUSION: Prestorage white cell reduction in apheresis components resulted in WBC reduction by several log10 with no evident adverse effect on platelet viability or function.

White cell-reduced platelet concentrates prepared by in-line filtration of platelet-rich plasma.

BACKGROUND: The importance of white cell (WBC) reduction in platelet concentrates (PCs) for component quality is undetermined. STUDY DESIGN AND METHODS: Eleven paired components, each derived from one of two whole-blood units given by a single donor on the same day, were studied. One PC was WBC reduced by filtration with an in-line, integral, prototype filter, and the other was produced from unfiltered platelet-rich plasma (PRP) by a standard method. In vitro tests performed on Day 1 and Day 5 were blood gases, plasma lactate, glucose, platelet ATP, mean platelet volume, morphology score, hypotonic stress ratio, extent of shape change in response to ADP, and beta-thromboglobulin. After 5 days of storage, each component pair was labeled with 51Cr or 111In and transfused for the estimation of percent recovery and survival. RESULTS: PCs using the in-line, prototypic filter had a platelet loss of approximately 15 percent and a variable 1 to 3 log10 reduction (average, 95%) in WBC content. The variation in filter WBC removal was related to PRP WBC content and indicated that the filter did not have the capacity for a 3 to 4 log10 removal when PRP WBC content exceeded 1 x 10(8). The in vitro and in vivo measures of platelet quality showed no meaningful differences between filtered and unfiltered PCs by paired t test. The mean differences in posttransfusion percent recoveries and survivals were 0.9 +/- 2.9 percent and 4 +/- 13 hours, respectively. Additional studies were performed using a larger filter with improved capacity. Those studies (n = 18) showed a significant improvement in filtration time and platelet yield and a consistent 3 to 4 log10 reduction in WBCs. Filtration time was 6.6 +/- 2.7 minutes, total PC WBCs were 9.6 +/- 4.6 x 10(4), and total PC platelets were 7.8 +/- 1.8 x 10(10) (mean +/- SD). CONCLUSION: Prestorage filtration of PRP and the preparation of filtered platelets do not result in any significant beneficial or adverse effect on subsequent platelet quality. With the large-capacity filter, consistent WBC reduction and good platelet yields are achieved.

The quality of red blood cells.

The evolving practice of medicine has required a number of changes in red cell product manufacture to ensure that the final product is more specifically tailored to the needs of the individual patient. As a result of the increasing concern over the risks of transfusion pharmaceutical standards of manufacture are now applied to blood component preparation. Studies have been undertaken to define the optimum method of blood processing, and newer technologies are emerging to allow acquisition of a more consistent dose of red cells in a fashion which may minimize the lesion of collection. Use of high efficiency 3+ generation filter technologies reduces leukokine build up during storage and improves the quality and purity of the stored blood product. The combination of new plasticizers for packaging and improved red cell additive solutions should allow the blood center to supply a more functional red cell with longer storage shelf life. Overall these developments should result in the provision of a more consistent dose of fully functional red cells to the recipient who will be less exposed to the undesirable sequelae of transfusion than previously.

Pseudothrombocytopenia in plateletpheresis donors.

BACKGROUND: EDTA pseudothrombocytopenia (PTCP) is an in vitro artifact in which the anticoagulation of blood with EDTA is associated with in vitro agglutination of platelets, resulting in a spuriously low platelet count. In apheresis donors, whole-blood samples for complete blood counts are routinely drawn into tubes anti-coagulated with EDTA. STUDY DESIGN AND METHODS: Records of apheresis donors were examined to identify persons in whom the postdonation counts were less than 100 x 10(9) per L. Identified donors were studied to confirm the presence of PTCP by drawing blood samples into EDTA, heparin, and trisodium citrate for serial platelet counts at room-temperature incubation. Platelet counts in citrated plasma were measured before and after the addition of EDTA. A single HLA-matched component from an identified PTCP donor was monitored for response by corrected count increment in the recipient. RESULTS: A total of nine donations were identified, involving 2 donors from a population of 945 donors (prevalence 0.2%). On testing, both donors were confirmed to have PTCP. The addition of EDTA to citrated plasma did not affect the platelet count. Response in a recipient to an HLA-matched component showed an acceptable corrected count increment. CONCLUSION: PTCP may occur in plateletpheresis donors and result in needless medical referral or donor deferral. PTCP does not appear to alter the yield content of the component or to be passively transferred to a recipient.

Characteristics of white cell-reduced red cells stored in tri-(2-ethylhexyl)trimellitate plastic.

BACKGROUND: Standard blood storage containers contain extractable plasticizers that accumulate in blood during storage and are an unintended transfusion product. However, extractable plasticizers have a protective effect on the red cell membrane and improve red cell storage variables. Prestorage white cell reduction also improves selected red cell storage variables. STUDY DESIGN AND METHODS: The study evaluated whether the beneficial effect of prestorage white cell reduction would offset the negative effect of the absence of extractable plasticizer in red cells stored in AS-3 for 42 days at 4 degrees C. Filtered red cells stored in polyvinylchloride containers with the nonextracting plasticizer, tri-(2-ethylhexyl)trimellitate (TEHTM), were compared to unfiltered red cells stored in polyvinylchloride containers with the extractable plasticizer di-(2-ethylhexyl)phthalate (DEHP). RESULTS: Poststorage supernatant potassium and red cell osmotic fragility were significantly higher in white cell-reduced TEHTM units than in unfiltered DEHP units. The mean 24-hour recovery of the filtered TEHTM red cells was significantly lower than that of the unfiltered DEHP red cells (69.1 +/- 7.4% vs. 77.1 +/- 5.1%, p < 0.05, n = 8). CONCLUSION: These data demonstrate that white cell reduction before 42-day storage in TEHTM containers with currently approved preservatives does not yield an acceptable red cell component.

Effects of 3-5 log10 pre-storage leucocyte depletion on red cell storage and metabolism.

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.

Effect on platelet properties of exposure to temperatures below 20 degrees C for short periods during storage at 20 to 24 degrees C.

BACKGROUND: When platelet concentrates (PCs) are shipped over long distances, it is not always possible to ensure that their temperature is maintained at 20 to 24 degrees C. In addition, PCs are not agitated as during routine storage. STUDY DESIGN AND METHODS: Studies have been conducted to evaluate how exposure to temperatures below 20 degrees C in the absence of agitation influences properties of platelets. In initial studies, exposure to 4 degrees C for 3 or 5 hours or to 12 degrees C for 5 or 17 hours on Day 2 of a 5- to 6-day storage period was associated with a loss of discoid shape. This was reflected by slightly lower but statistically different morphology scores after storage compared to those observed with control platelets that were stored only at 20 to 24 degrees C. In addition, a qualitative difference in morphology was noted in controls and PCs held at 16 degrees C for 17 hours. In more detailed studies, both the in vivo viability and in vitro properties of platelets exposed between Day 1 and Day 2 to either 12 degrees C or 16 degrees C for 17 hours were evaluated. The protocol involved a paired study design (n = 4 for each exposure temperature) with the simultaneous storage of two identical PCs, one exposed to 12 or 16 degrees C and the other one maintained at 20 to 24 degrees C throughout the 5-day storage. RESULTS: Exposure to 12 degrees C significantly reduced (p < 0.05 by paired t test) the in vivo recovery to 37.6 +/- 13.8 percent (mean +/- 1 SD) from 47.8 +/- 11.5 percent and the survival time to 2.0 +/- 0.3 days from 6.5 +/- 1.4 days. On exposure to 16 degrees C, the differences in viability from those of control units were much less but still significant. The in vivo recovery was 42.7 +/- 3.8 percent compared to 49.2 +/- 3.0 percent and the survival time was 3.5 +/- 1.2 days compared to 6.6 +/- 0.3 days. The loss of in vivo viability of the test platelets was associated with a loss of discoid shape, as reflected by morphology scores, extent of shape change, and mean platelet volume. In addition, platelet metabolism also appeared to be affected, as suggested by increased lactate production. All of the in vitro properties except for total ATP and residual glucose that were statistically different from those of controls on exposure to 12 degrees C were also significantly different on exposure to 16 degrees C. CONCLUSION: These findings demonstrate that platelets undergo substantial changes in in vivo viability and in vitro properties when they are exposed to temperatures below 20 degrees C for short periods.

Sustained elevation of intracellular cyclic 3'-5' adenosine monophosphate is necessary for preservation of platelet integrity during long-term storage at 22 degrees C.

Preservation of platelet integrity and responsiveness was examined in platelet concentrates prepared in the presence of various formulations and combinations of platelet-activation inhibitors affecting intracellular levels of cyclic 3'-5' adenosine monophosphate (cAMP). Platelet concentrates were prepared and stored in an artificial medium for two weeks at 22 degrees C. Markers of metabolic activity (pH, lactate, pO2, pCO2 in the medium), aggregation response, hypotonic shock response, and glycoprotein Ib (GPIb) expression were assessed along with direct measurements of cAMP in platelet pellets and thromboxane B2 (TxB2) in the supernate. The platelet concentrates prepared with only adenylate-cyclase stimulators (prostaglandin E-1 or forskolin) showed less maintenance of the integrity and responsiveness markers and greater loss of GPIb than concentrates prepared with phosphodiesterase inhibitors (theophylline or caffeine) or combinations with the above. These results were correlated with the ability of these compounds to sustain elevation of cAMP above basal level during the entire extended-storage period. The strong correlation (rs = -0.67) between elevation of cAMP levels and suppression of TxB2 production suggests that the phosphodiesterase inhibitors provided better protection than stimulators of adenylate cyclase alone through a reduction in platelet activation and its deleterious effects on preservation of platelets during storage.

Evaluation of apheresis platelet concentrates collected with a reduced (30-ml) collection chamber with resuspension and storage in a synthetic medium.

Recently, the CS-3000 Plus Blood Cell Separator with the TNX-6 platelet separation chamber insert has been furnished with a small-volume (30-ml) collection chamber. In this study, a platelet synthetic medium containing glucose and bicarbonate (PSM) was used for resuspension and storage of this highly concentrated platelet product. Eighteen donors participated in a paired study design where each participant donated platelets on two occasions, once following collection in a standard chamber with resuspension and storage in plasma and once following collection in the new chamber with resuspension and storage in PSM. Substantially higher total platelet counts were obtained using platelets collected in the small chamber and stored in PSM as compared to control (4.4 +/- 0.9 x 10(11) vs. 3.5 +/- 0.9 x 10(11) platelets, p < 0.01 by paired t test). After 5 days of storage, PSM-stored platelets demonstrated higher ATP levels, less lactate dehydrogenase in the supernatant and increased lactate production with resulting lower pH at day 5 of storage (6.94 +/- 0.15 vs. 7.08 +/- 0.09, p < 0.05). There were no statistically significant differences of the survival by multiple-hit estimation of PSM-stored as compared to plasma-stored platelets as determined by 111In labeling and infusion. A slight decrease in the initial percent recovery with the additive-suspended as compared to suspended plasma cells was noted: 50 +/- 8 versus 54 +/- 9%, respectively (p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Evaluation of platelet concentrates stored for 5 days with reduced plasma volume.

BACKGROUND: Currently, platelet concentrates (PCs) are stored in a suspending plasma volume of 45 to 65 mL. Previous studies using second-generation containers indicated that PCs stored for 5 days at volumes less than 30 mL have reduced in vivo percentage recoveries as compared to PCs stored at volumes of 50 mL or more. STUDY DESIGN AND METHODS: This study has evaluated the effect of PC plasma volume on the maintenance of in vivo and in vitro platelet properties following 5 days of storage, with the purpose of establishing the minimum plasma volume in the range of 30 to 50 mL. Twenty paired studies were performed in which identical populations of platelets from the same donor (obtained by double manual apheresis) were stored in a normal volume (55-60 mL, control) and reduced volume (30-50 mL, test) of plasma. Comparison of in vivo viability between test and control PCs was performed after random radiolabeling of 1 unit with 51Cr and of the other with 111In, with simultaneous transfusion and with calculation of percentage recovery and the area below the survival curve (integral) as measures of viability. RESULTS: When test unit volumes were > or = 35 mL, essentially identical platelet survival curves and in vitro results were obtained for test and control. The integral and the percentage recovery for the test units were (mean, 95% confidence interval) 98.7 (96.3-101.0) and 99.0 percent (94.7-103.3) of those values in the control units, respectively. Test units with volume < or = 34 mL demonstrated reduced in vivo viability with integral and percentage recovery of 81.1 (68.9-93.3) and 80.4 (69.3-91.5), respectively, as compared to the control units. This loss was associated with increased metabolic activity (lactate production), which may suggest platelet activation due to the increased surface-to-PC volume ratio. CONCLUSION: These results show that the storage volume of PCs may be reduced from 50 to 60 mL to 35 to 40 mL without any significant decrease in in vivo or in vitro platelet quality.

Storage of pools of six and eight platelet concentrates.

Platelet concentrates (PCs) prepared from units of whole blood are routinely stored singly at 20 to 24 degrees C and pooled prior to transfusion. Studies have been conducted to evaluate the in vitro properties of pools of six (n = 19) and eight (n = 17) ABO-identical PCs after storage, with comparative studies involving single units (n = 33). The pools were prepared using the sterile connecting device. One-day-old and 3-day-old PCs were pooled and stored for a total of 5 days in a container system consisting of two 1000-mL polyolefin containers. The pooled platelet suspension was divided approximately equally between the two containers. The platelet count was reduced by less than 5 percent during storage of the pools, which is similar to the reduction found with storage of control units of single PCs. The volume loss due to pooling was 9.6 +/- 1.9 percent (mean +/- 1 SD). The pH of the PC pools was approximately 7.0 after 5 days of storage, with no pool having a pH below 6.2. In vitro platelet properties, such as morphology score, extent of shape change induced by ADP, total ATP, aggregation response to ADP and collagen, response to hypotonic stress, lactate dehydrogenase discharge, and beta-thromboglobulin release, were similar for pools and control single PCs. In addition, comparable low levels of thymidine uptake were detected in the mononuclear leukocyte fraction of pooled and unpooled PCs that were stored for 5 days at 20 to 24 degrees C, which indicates that the mixing of lymphocytes in the pool did not stimulate in vitro immunologic reactions.(ABSTRACT TRUNCATED AT 250 WORDS)