Is hydroxyethyl starch necessary for sedimentation of bone marrow?

Hydroxyethyl starch (HES) is used to separate hematopoietic progenitor cells after bone marrow (BM) collection from red blood cells. The aims were to study alternatives for HAES-steril (200 kDa; not available anymore) and to optimize the sedimentation process. Using WBC-enriched product (10 × 10(9) WBC/L), instead of BM, sedimentation at 10% hematocrit using final 0.6 or 0.39% Voluven (130 kDa) or without HES appeared to be good alternatives for 0.6% HAES-steril. MNC recovery >80% and RBC depletion >90% was reached. Optimal sedimentation was reached using 110-140 mL volume. Centrifugation appeared not suitable for sedimentation. Additional testing with BM might be necessary to confirm these results.

Comparison of cryopreservation bags for hematopoietic progenitor cells using a WBC-enriched product.

Hematopoietic progenitor cells (HPC) are stored in cryopreservation bags that are resistant to liquid nitrogen. Since Cryocyte bags of Baxter (B-bags) are no longer available, an alternative bag was sought. Also, the influence of freezing volume was studied. Miltenyi Biotec (MB)- and MacoPharma (MP)-bags passed the integrity tests without failure. Comparing MB- and MP-bags with B-bags, no difference in WBC recovery or viability was found when using a WBC-enriched product as a "dummy" HPC product. Further, a freezing volume of 30 mL resulted in better WBC recovery and viability than 60 mL. Additonal studies using real HPC might be necessary.

Optimization of the freezing process for hematopoietic progenitor cells: effect of precooling, initial dimethyl sulfoxide concentration, freezing program, and storage in vapor-phase or liquid nitrogen on in vitro white blood cell quality.

BACKGROUND: Adding dimethyl sulfoxide (DMSO) to hematopoietic progenitor cells (HPCs) causes an exothermic reaction, potentially affecting their viability. The freezing method might also influence this. The aim was to investigate the effect of 1) precooling of DMSO and plasma (D/P) and white blood cell (WBC)-enriched product, 2) DMSO concentration of D/P, 3) freezing program, and 4) storage method on WBC quality. STUDY DESIGN AND METHODS: WBC-enriched product without CD34+ cells was used instead of HPCs. This was divided into six or eight portions. D/P (20 or 50%; precooled or room temperature [RT]) was added to the WBC-enriched product (precooled or RT), resulting in 10% DMSO, while monitoring temperature. The product was frozen using controlled-rate freezing ("fast-rate" or "slow-rate") and placed in vapor-phase or liquid nitrogen. After thawing, WBC recovery and viability were determined. RESULTS: Temperature increased most for precooled D/P to precooled WBC-enriched product, without influence of 20 or 50% D/P, but remained for all variations below 30°C. WBC recovery for both freezing programs was more than 95%. Recovery of WBC viability was higher for slow-rate freezing compared to fast-rate freezing (74% vs. 61%; p < 0.05) and also for 50% compared to 20% D/P (two test variations). Effect of precooling D/P or WBC-enriched product and of storage in vapor-phase or liquid nitrogen was marginal. CONCLUSION: Based on these results, precooling is not necessary. Fifty percent D/P is preferred over 20% D/P. Slow-rate freezing is preferred over fast-rate freezing. For safety reasons storage in vapor-phase nitrogen is preferred over storage in liquid nitrogen. Additional testing using real HPCs might be necessary.

Noninvasive pH monitoring of platelet concentrates: a large field test.

BACKGROUND: Developing new quality control methods for platelet concentrates (PCs) can contribute to increasing transfusion safety and efficiency. The aim of this study was to investigate in a large field test the quality of expired PCs and whether 100% noninvasive pH monitoring can be used to predict PC quality. STUDY DESIGN AND METHODS: The pH of 13,693 PCs produced for transfusion was monitored daily using Blood Storage, Inc.'s pH sterile, automated fluoroscopic evaluation technology. Upon indication of compromised quality or expiration, PCs were returned and in vitro tests were performed. RESULTS: A total of 998 PCs were returned, of which 962 outdated, 26 had a positive BacT/ALERT reaction, seven had aggregates, one was without swirl, one had low pH, and one had high pH. BacT/ALERT was faster in identifying bacterial contamination than pH measurements. The pH at the end of the storage period was significantly lower than at the beginning. In vitro tests indicated that while the PC quality was acceptable upon expiration, it rapidly declined after expiration. CONCLUSION: In this setting where the vast majority of PCs were of good quality and within acceptable pH limits, daily, noninvasive routine pH measurement has limited added value in identifying quality-compromised PCs.

Hemolysis of red blood cells during processing and storage.

BACKGROUND: During processing and storage, red blood cells (RBCs) undergo changes and cell injury resulting in hemolysis. Mostly, the separation of whole blood in top-and-bottom quadruple bag systems with break openings takes less than 4 minutes. However, longer separation times are not uncommon. The aims were to investigate whether hemolysis is increased in RBCs with longer separation time (RBCs(>6 min)) compared to regular RBCs (RBCs(reg)), to measure hemolysis increase during storage and to study whether frequency of hemolytic donations is donor dependent. STUDY DESIGN AND METHODS: RBCs(>6 min) (n = 172) and 172 matched controls were tested for hemolysis on Days 1 and 21 RBC units from each group were stored at 4 ± 2°C and tested again after 5 weeks. Donor dependency was retrospectively investigated for 100 hemolytic RBC units. RESULTS: RBCs(>6 min) exhibited a higher mean hemolysis rate than RBCs(reg) (0.058% vs. 0.033%). Four RBC units were hemolytic (>0.8%), all RBCs(>6 min) (2.36%). During storage, hemolysis in both groups increased with 0.24%. Hemolysis frequency did not seem to be donor dependent. CONCLUSIONS: Increased separation time is a useful screening tool for potentially increased hemolysis rate in RBCs. Hemolysis rate increased during storage equally in both groups. Hemolysis frequency appears donor independent.

Allogeneic single-donor cryoseal produced from fresh-frozen quarantine apheresis plasma as alternative for multidonor or autologous fibrin sealants.

BACKGROUND: Fibrin sealant is a human blood product consisting of two components: cryoprecipitate and thrombin. Commercial fibrin sealants are produced from multidonors, increasing the viral risk, and contain fibrinolytic inhibitors such as tranexamic acid or bovine aprotinin. Autologous fibrin sealants reduce the viral risk and are mostly produced during a surgical procedure or well in advance. Alternatively, the allogeneic single-donor fibrin sealant cryoseal can be used. In this study cryoseal was characterized and the manufacturing consistency of the production process was investigated. STUDY DESIGN AND METHODS: Cryoseal was produced from plasma collected on apheresis machines using a commercial device. In a research setting the protein composition and recovery were determined. Also, the manufacturing consistency of the production process was tested in a research setting as well as in a routine setting. RESULTS: In the research setting all produced cryoseal met the quality control requirements of a clotting time of less than 10 seconds and the presence of Factor (F)XIII (qualitative). In the routine setting, one procedure per year did not meet these requirements. The protein composition showed the following mean ± standard deviation (%recovery) results: thrombin 25.7 ± 11.1 IU/mL, fibrinogen 19.9 ± 4.6 (15%) mg/mL, FVIII 15.6 ± 5.4 (44%) IU/mL, FXIII 2.7 ± 0.7 (6%) IU/mL, and plasminogen 1.8 ± 0.2 (4%) U/mL. In both research and routine settings the production process resulted in a consistent product. CONCLUSION: The cryoseal manufacturing process resulted in a consistent product, which meets the predetermined specifications. The single-donor origin and the absence of fibrinolytic inhibitors make cryoseal a good alternative for multidonor and autologous fibrin sealants.

Evaluation of overnight hold of whole blood at room temperature before component processing: effect of red blood cell (RBC) additive solutions on in vitro RBC measures.

BACKGROUND: Whole blood (WB) can be held at room temperature (18-25°C) up to 8 hours after collection; thereafter the unit must be refrigerated, rendering it unsuitable for platelet (PLT) production. Overnight hold at room temperature before processing has logistic advantages, and we evaluated this process in an international multicenter study for both buffy coat (BC)- and PLT-rich plasma (PRP)-based blood components and compared three red blood cell (RBC) additive solutions (ASs) for their ability to offset effects of overnight hold. STUDY DESIGN AND METHODS: Nine centers participated; seven used the BC method, and two used the PRP method. Four WB units were pooled and split; 1 unit was processed less than 8 hours from collection (Group A), and the other three (Groups B, C, and D) were held at room temperature and processed after 24 to 26 hours. RBCs in Groups A and B were resuspended in saline-adenine-glucose-mannitol, Group C in phosphate-adenine-guanosine-glucose-saline-mannitol, and Group D in ErythroSol-4 RBCs were stored at 2 to 6°C for 49 days. RESULTS: RBCs from overnight-held WB had lower 2,3-diphosphoglycerate (2,3-DPG) and higher adenosine triphosphate (ATP). At the end of storage there were no differences between groups, apart from a slightly higher hemolysis in Group B. ErythroSol-4 showed a slightly higher initial ATP and 2,3-DPG content, but at the end of storage no differences were found. CONCLUSION: Overnight hold of WB before processing has no lasting deleterious effects on in vitro quality of subsequently prepared components. The use of different RBC ASs did not appear to offer significant advantages in terms of RBC quality at the end, regardless of the processing method.

Is red blood cell rheology preserved during routine blood bank storage?

BACKGROUND: Red blood cell (RBC) units stored for more than 2 weeks at 4 degrees C are currently considered of impaired quality. This opinion has primarily been based on altered RBC rheologic properties (i.e., enhanced aggregability, reduced deformability, and elevated endothelial cell interaction), during prolonged storage of nonleukoreduced RBC units. In this study, the rheologic properties and cell variables of leukoreduced RBC units, during routine blood bank storage in saline-adenine-glucose-mannitol, were investigated. STUDY DESIGN AND METHODS: Ten leukoreduced RBC units were stored at the blood bank for 7 weeks at 4 degrees C. RBCs were tested weekly for aggregability, deformability, and other relevant variables. RESULTS: RBC aggregability was significantly reduced after the first week of storage but recovered during the following weeks. After 7 weeks aggregability was slightly, but significantly, reduced (46.9 + or - 2.4-44.3 + or - 2.2 aggregation index). During storage the osmotic fragility was not significantly enhanced (0.47 + or - 0.01% phosphate-buffered saline) and the deformability at shear stress of 3.9 Pa was not significantly reduced (0.36 + or - 0.01 elongation index [EI]). The deformability at 50 Pa was reduced (0.58 + or - 0.01-0.54 + or - 0.01 EI) but remained within reference values (0.53 + or - 0.04). During 5 weeks of storage, adenosine triphosphate was reduced by 54% whereas mean cell volume, pH, and mean cell hemoglobin concentration were minimally affected. CONCLUSIONS: RBC biochemical and physical alterations during storage minimally affected the RBC ability to aggregate and deform, even after prolonged storage. The rheologic properties of leukoreduced RBC units were well preserved during 7 weeks of routine blood bank storage.

Counting platelets in platelet concentrates on hematology analyzers: a multicenter comparative study.

BACKGROUND: Hematology analyzers are designed to count whole blood samples, but are also used by blood centers to perform quality control on blood components. In platelet (PLT) concentrates, the number of PLTs is approximately fivefold higher and red blood cells are absent, causing variable PLT counting results. It was our aim to compare currently used hematology analyzers for counting PLTs in PLT concentrates using fixed human PLTs. STUDY DESIGN AND METHODS: PLT samples were fixed, diluted into seven concentration levels (plus one blank), aliquoted, and shipped to 68 centers. Evaluable data were obtained for 89 hematology analyzers. All samples were counted six times, and results were reported to the coordinating center. The overall group mean was calculated, and the percentage deviation from this mean was calculated for each analyzer. RESULTS: At PLT levels relevant for blood centers, 750 x 10(9) to 2000 x 10(9) per L, analyzers gave results that were between 35 percent lower and 16 percent higher than the overall group mean. Within a group of analyzers, results were comparable with coefficient of variations usually below 10 percent, indicating that the observed differences were caused by instrument characteristics. A smaller study with fresh, unfixed PLT samples showed that analyzers behaved similarly for fixed and fresh PLTs. CONCLUSION: With a wide array of currently used hematology analyzers, a marked difference was determined for the PLT counts of fixed human-based identical samples provided to 68 laboratories by a centralized facility. A gold standard method is needed to allow for more valid interlaboratory comparisons between hematology analyzers.

Platelet capacity of various platelet pooling systems for buffy coat-derived platelet concentrates.

BACKGROUND: Platelet (PLT) storage lesions might depend on the total PLT count in the storage container and also on the PLT pooling system, especially the storage container, that is used for preparation of PLT concentrates (PCs). In this study, the PLT capacity of four commercially available PLT pooling systems was studied. MATERIALS AND METHODS: Four PCs were prepared in pooling systems of Baxter, Fresenius, Terumo, or Pall. The PCs were pooled and divided with various total PLT counts over the four storage containers (<225 x 10(9), 225 x 10(9)-324 x 10(9), 325 x 10(9)-424 x 10(9), and >424 x 10(9) PLTs). Volumes were kept equal by adding plasma to PCs with less than 425 x 10(9) PLTs until a same volume as for PCs with more than 424 x 10(9) PLTs was reached. PCs were stored at room temperature and tested for various in vitro variables on Days 1, 3, 5, 7, and 9. Paired experiments were repeated for each system five times. RESULTS: In vitro variables remained good for 9 days, that is, swirling score of 2 or more, pH value of 6.8 or more, glucose level of 10 mmol per L or more, lactate level of less than 25 mmol per L, and CD62p expression of less than 50 percent, for PCs in Baxter systems with more than 225 x 10(9) PLTs, for PCs in Fresenius and Terumo systems with 225 x 10(9) to 424 x 10(9) PLTs, and for PCs in Pall systems with fewer than 425 x 10(9) PLTs. CONCLUSION: PLT capacity depended on the PLT pooling systems used. All systems provide acceptable storage conditions. The Baxter system was the only system with capacity for more than 424 x 10(9) PLTs per PC.

Overnight or fresh buffy coat-derived platelet concentrates prepared with various platelet pooling systems.

BACKGROUND: For logistic reasons, possibilities to produce both platelet (PLT) concentrates prepared from fresh or overnight-stored whole blood (fresh and o/n PCs, respectively) are convenient. The consequences of both possibilities are not well described. The PLT pooling system used might also influence the condition of PCs. Our aim was to compare fresh and o/n PCs with different PLT pooling systems. STUDY DESIGN AND METHODS: Fresh and o/n PCs were prepared from buffy coats and plasma in PLT pooling systems of Baxter, Fresenius, Terumo, or Pall (n = 5). PCs were stored for 9 days. The in vitro quality was determined by the PLT count, pH, glucose, lactate, pO(2), pCO(2), CD62P expression, and annexin V binding. RESULTS: The o/n PCs showed higher PLT count (approx. 460 x 10(9)/PC vs. approx. 310 x 10(9)/PC), pCO(2), and lactate concentration and lower pH, pO(2), glucose concentration, CD62P expression (until Day 5), and annexin V binding (until Day 7) compared with fresh PCs (p < 0.05). Only for o/n PCs in the Baxter and Fresenius systems did the pH and glucose concentration remain higher, and the lactate concentration and CD62P expression remained lower than that of o/n PCs in the Pall and Terumo systems. The pH for fresh PCs in the Baxter and Fresenius systems was more often greater than 7.4 than for fresh PCs in the Terumo or Pall systems. CONCLUSION: The quality of PCs depended on whether PCs were prepared from fresh or overnight-stored whole blood and on the used PLT pooling system. The main difference between fresh and o/n PCs was the PLT count.

Flow cytometric measurement of CD62P (P-selectin) expression on platelets: a multicenter optimization and standardization effort.

BACKGROUND: One of the variables to determine the quality of platelets (PLTs) in vitro is measurement of CD62P expression. Different protocols are in use, however, making comparison of results virtually impossible. It was our aim to develop a uniform CD62P protocol that would yield comparable results in various laboratories. STUDY DESIGN AND METHODS: The effects of fixation, source and dilution of CD62P antibody, source of immunoglobulin G (IgG) isotypic antibody, and analysis of results were investigated. Once the optimal variables were defined, comparative studies were performed at five participating centers. In the final comparative study, eight split PLT concentrates were shipped to the centers, where samples were stained and fixed according to the uniform protocol. Analyses were performed using commercially available flow cytometers (BD Biosciences and Beckman Coulter). RESULTS: Uniformity between centers could be achieved by using a single clone for CD62P and IgG monoclonal antibody. A protocol was selected using fixation with 0.5 percent methanol-free formaldehyde. To increase conformity between flow cytometers, in the analysis of electronic data the thresholds of the isotypic control were set at 0.5 percent for the BD Biosciences and 2 percent for the Beckman Coulter flow cytometers. In the final comparative study, the 95 percent confidence intervals (CIs) for CD62P ranged between 8 and 21 percent in fresh and 20 to 40 percent in 8-day-old PLT concentrates. CONCLUSION: A uniform CD62P staining protocol and subsequent analysis can be used at multiple centers using different flow cytometers, yielding comparable results with acceptable 95 percent CIs.

Platelet counting in platelet concentrates with various automated hematology analyzers.

BACKGROUND: Hematology analyzers use impedance, optical, and/or immunologic techniques for counting platelets (PLTs). PLT counting in whole blood has been validated thoroughly; however, this is not the case for PLT counting in PLT concentrates (PCs), in which red cells (RBCs) are absent. Therefore, this study is focused on PLT counting in PCs to study use of ethylenediaminetetraacetate (EDTA), carryover, and accuracy of the analyzers. STUDY DESIGN AND METHODS: In total six hematology analyzers (AcT 8, Beckman Coulter; ADVIA 2,120, Bayer; Cell-Dyn 4,000, Abbott; Onyx, Beckman Coulter; K4,500, Sysmex; and XT 2,000i, Sysmex) were tested for PLT counting. PC samples with various PLT concentrations were made (0-1,700 x 10(9)/L) and measured 10 times. Carryover was determined five times. RESULTS: PC samples (1,000 x 10(9) PLTs/L) in EDTA tubes showed significantly higher PLT counts than samples in "dry" tubes for all analyzers except for the Cell-Dyn 4,000 with the impedance technique. Carryover was not more than 0.3 percent for all analyzers. The K4,500 showed the most accurate results, whereas the Cell-Dyn 4,000 with the impedance technique had low accuracy due to an overestimation of more than 20 percent. CONCLUSION: Most tested analyzers seemed to be suitable for counting PLTs in PCs. All hematology analyzers should be validated for counting PLTs in absence of RBCs as is the case in PCs, in addition to validation of PLT counting in whole blood.

Interruption of agitation of platelet concentrates: a multicenter in vitro study by the BEST Collaborative on the effects of shipping platelets.

BACKGROUND: Transported platelets (PLTs) are not under continuous agitation. The aim of this study was to determine whether PLTs shipped between 24 and 48 hours would be able to maintain a pH(22 degrees C) value of 6.5 at the end of 7 days of storage. STUDY DESIGN AND METHODS: Six laboratories prepared leukoreduced PLTs. PLT pools were divided into low and high PLT concentration with paired shipped (20-43 hr) and unshipped controls. Units were under continuous agitation at 22 +/- 2 degrees C when not being transported. In vitro measures including pH, pO(2), and pCO(2) were determined over 7 days. RESULTS: Ninety-two PLT components from 24 pools were eligible for analysis. One unshipped control and three shipped products failed to maintain a pH(22 degrees C) value of 6.5 through 7 days. In vitro characteristics were maintained slightly better over 7 days of storage in the unshipped control arms. PLT concentration, shipping time, and their interaction were significant determinants of the final pH at the end of storage (p < 0.05). Lactate generation rate increased by 35 +/- 2 (mean +/- SE) micromol per 10(12) PLTs per hour over baseline during shipping (p < 0.0001). After restoration of standard blood banking conditions with agitation, this rate dropped 24 +/- 2 micromol per 10(12) PLTs per hour (p < 0.0001). CONCLUSION: PLTs in plasma shipped for at least 20 to 24 hours maintain a pH(22 degrees C) value of 6.5 for 7 days. A longer shipping time may result in a pH(22 degrees C) value of 6.5. During shipping, glycolysis was up regulated in these PLTs resulting in increased lactic acid production. After restoration of agitation, shipped products down regulated glycolysis. These effects should be accounted for in the development of PLT storage and transportation systems.

The effect of whole-blood storage time on the number of white cells and platelets in whole blood and in white cell-reduced red cells.

BACKGROUND: Whole blood (WB) can be stored for some time before it is processed into components. After introduction of universal white cell (WBC) reduction, it was observed that longer WB storage was associated with more residual WBCs in the WBC-reduced red cells (RBCs). Also, weak propidium iodide (PI)-positive events were observed in the flow cytometric WBC counting method, presumably WBC fragments. The effect of storage time on the composition of WB and subsequently prepared WBC-reduced RBCs was studied. STUDY DESIGN AND METHODS: WB was collected in bottom-and-top collection systems with inline filters, obtained from Baxter, Fresenius, or MacoPharma. Units were stored at room temperature and separated into components in 4-hour intervals between 4 and 24 hours after collection. RBCs were WBC-reduced by inline filtration (approx. 50/group). RESULTS: Platelet (PLT) counts were lower in WB stored for 4 to 8 hours compared to 20 to 24 hours (mean +/- SD): 79 +/- 31 versus 102 +/- 30 for Baxter (p < 0.01); 91 +/- 31 versus 101 +/- 35 for Fresenius (not significant); and 73 +/- 47 versus 97 +/- 31 (all x 10(9) per unit) for MacoPharma (p < 0.01), respectively. The median residual WBC counts in WBC-reduced RBCs for WB stored for 4 to 8 and 20 to 24 hours were 0.03 versus 0.17 for Baxter (p < 0.001), 0.00 versus 0.06 for Fresenius (p < 0.001), and 0.13 versus 0.26 (all x 10(6) per unit) for MacoPharma (not significant), respectively. All WBC-reduced RBCs contained fewer than 5 x 10(6) WBCs per unit. A longer storage time of WB was associated with more weak PI-positive events, irrespective of the filter. CONCLUSION: Longer storage of WB before processing results in counting higher numbers of PLTs in WB, higher numbers of WBCs in WBC-reduced RBCs, and more weak PI-positive events.

Storage of platelets in additive solutions: effects of magnesium and/or potassium.

BACKGROUND: Previous studies indicate that platelet concentrates (PCs) in a platelet additive solution (PAS) containing citrate, acetate, and sodium chloride (PAS-2) show a significantly higher increase of CD62+ platelets than PCs in other brands of PAS containing Mg(2+) and K(+). To investigate whether this difference can be explained by the presence of Mg(2+) and/or K(+) in the storage medium, we performed paired studies comparing storage of PCs in PAS-2 to PAS-2 with either Mg(2+) or K(+) or both in combination. STUDY DESIGN AND METHODS: PCs from pooled buffy coats were prepared in either PAS-2 or PAS-2 with Mg(2+) or K(+) or both in combination (PAS-2 modified). Different volumes of MgCl(2) solution (1 mol/L) and/or KCl solution (1 mol/L) were added to PAS-2 to obtain various concentrations. After preparation and during storage (at Days 3 and 7), pH, pCO(2), pO(2), HCO(3)(-), and CD62 (%) were measured. RESULTS: During 7 days of storage, pH was very stable (6.9-7.2) in all PCs. At Day 7, platelet CD62 expression was 49 percent (PAS-2), 41 percent (PAS-2 with 1.5 mmol/L Mg(2+)), and 38 percent (PAS-2 with 4.5 mmol/L Mg(2+)). With added K(+), at Day 7, expression of CD62 was 55 percent (PAS-2), 39 percent (PAS-2 with 4.5 mmol/L K(+)), and 35 percent (PAS-2 with 9.0 mmol/L K(+)). In PAS-2 modified (PAS-2 with 1.5 mmol/L Mg(2+) and 4.5 mmol/L K(+)) and CPD plasma, the corresponding CD62 values were 23 and 35 percent, respectively. CONCLUSION: The combination of Mg(2+) and K(+) gave significantly (p < 0.05) lower platelet CD62 expression in the storage medium than in PAS-2. The effects of these differences on platelet metabolism and in vivo properties remain to be investigated.

Multicenter evaluation of two flow cytometric methods for counting low levels of white blood cells.

BACKGROUND: Flow cytometric methods can be used to count residual white blood cells (WBCs) in WBC-reduced blood products, which should contain fewer than 1 x 10(6) WBCs per unit (approximately 3.3 WBCs/ microL). In this study two flow cytometric methods for counting WBCs under routine conditions in nine laboratories were evaluated. STUDY DESIGN AND METHODS: Panels of red blood cells (RBCs), platelets (PLTs), and plasma were prepared containing 33.3, 10.0, 3.3, 1.0, and 0.3 WBCs per microL and counted with flow cytometric methods (either LeucoCOUNT, BD Biosciences, four laboratories; or LeukoSure, Beckman Coulter, five laboratories). Requirements were that at the level of 3.3 WBCs per microL, coefficient of variation was < or =20 percent and accuracy was > or =80 percent. Routine flow cytometric quality control (QC) data of WBC-reduced blood products from two laboratories were analyzed. RESULTS: At the level of 3.3 WBCs per microL, none of the laboratories met the requirements for all three blood products. The LeucoCOUNT method met requirements at more laboratories than the LeukoSure method for RBCs and PLTs, but the opposite was true for plasma. Routine QC data showed that >99 percent of the flow cytometric measurements for WBC-reduced products was below the 95 percent prediction interval at 3.3 WBCs per microL. CONCLUSION: None of the laboratories met the requirements for accuracy and precision for all three blood products. Nevertheless, routine results showed that in >99 percent of the products, WBC counts were below guideline limits. Therefore, both flow cytometric methods are suitable for QC with pass-fail criterion.