Evaluation of a strategy to limit blood donor exposure in high risk premature newborns based on clinical estimation of transfusion need

OBJECTIVES: Reservation of dedicated series of pedipacks, consisting of 3 to 4 units of 70 ml filtered red cell concentrate in additive solution SAGM from 1 donor, may reduce donor exposure. In this prospective efficacy study the benefits, release and expiration of pedipacks (PP) assigned to preterm infants requiring neonatal intensive care are analyzed. METHODS: On the basis of clinical assessment of the need for multiple transfusions, 96 preterm neonates (gestational age < 32 wks and/or birth weight < 1500 g) were assigned to either the high risk group (HRG), who were to receive dedicated donor blood units, or the low risk group (LRG). Inclusion criteria for HRG were 1) estimated time of admission > 21 days and 2) expected need for multiple transfusions due to clinical cardiorespiratory instability, prolonged parental feeding or frequent blood sampling. To reduce wastage of donor blood, dedication of donor blood units was limited to 21 days. RESULTS: 50 series (192 PP) were assigned to 42 HRG infants. Two HRG infants received 3 series, 4 received 2 series and 36 received 1 series of PP. Mean transfusion rate was 3.1 PP in the HRG and 0.4 in the LRG. In the LRG 35 of 54 were not transfused, 19 received 1 to 2 PP. In both groups transfused newborns were exposed to 1.1 donors in average. In the HRG of 192 PP, 137 PP (71%) were used within 21 days, and another 30 (16%) before the expiration date < 35 days. Twenty five PP (13%) expired, mainly because of logistical problems in the introduction phase. CONCLUSION: Assignment of dedicated PP on the basis of clinical parameters at entry considerably reduces donor exposure in HRG. Wastage of dedicated blood transfusions was reduced by limitation of the dedicated period (21 days). In terms of efficacy, reservation and use of PP can be optimized by standardized administrative measures.     

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, pO₂, pCO₂, CD62P expression, and annexin V binding.
RESULTS: The o/n PCs showed higher PLT count (approx. 460 × 10⁹/PC vs. approx. 310 × 10⁹/PC), pCO₂, and lactate concentration and lower pH, pO₂, 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.     

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 × 10⁹, 225 × 10⁹‐324 × 10⁹, 325 × 10⁹‐424 × 10⁹, and >424 × 10⁹ PLTs). Volumes were kept equal by adding plasma to PCs with less than 425 × 10⁹ PLTs until a same volume as for PCs with more than 424 × 10⁹ 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 × 10⁹ PLTs, for PCs in Fresenius and Terumo systems with 225 × 10⁹ to 424 × 10⁹ PLTs, and for PCs in Pall systems with fewer than 425 × 10⁹ 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 × 10⁹ PLTs per PC.     

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.     

Comparison of five platforms for enumeration of residual leucocytes in leucoreduced blood components

The need for quality control of leucoreduction of blood products has led to the development of various methods to count low levels of residual leucocytes. We compared five platforms side-by-side: the Nageotte haemocytometer and four based on fluorescent staining of nuclei: two flowcytometers (Beckman Coulter, BD Biosciences) with methods based on counting beads, a volumetric flow cytometer (Partec) and the microvolumic fluorimeter ImagN2000 (BD Biosciences), all according to their manufacturers' recommended methods. Analysis of double-filtered red cell concentrates (RCCs) and platelet concentrates (PCs), spiked with various numbers of leucocytes, revealed good linearity for all methods over the range of 1.6-32.7 leucocytes/microl, all with r(2) > 0.99. At the rejection level of leucocyte-reduced blood components, i.e. 1 x 10(6) per unit corresponding with approximately 3.3 leucocytes/microl, the Nageotte haemocytometer had low accuracy (0% for RCCs, 56% for PCs), and was relatively imprecise [coefficient of variance (CV) of 34% and 30% respectively]. The Partec flow cytometer gave good results for RCCs (accuracy 67%, CV 22%), but not for PCs (accuracy 0%, CV 25%). The ImagN2000 had an accuracy of 44% for RCCs and 89% for PCs, but the precision was variable (CV 32% for RCCs, 15% for PCs). The best results were obtained with the Beckman Coulter (RCCs: accuracy 86%, CV 13%, PCs: accuracy 67%, CV 16%), and BD Biosciences platforms (RCCs: accuracy 100%, CV 10%; PCs: accuracy 89%, CV 11%). We conclude that, at the rejection level of 1 x 10(6) leucocytes per unit, the widely used Nageotte haemocytometer performs poorly in terms of inaccuracy and imprecision, and that both counting-bead-based, flow cytometric methods performed best.     

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.     

In vivo and in vitro comparison of platelets stored in either synthetic media or plasma

Since 1980, several synthetic media have been developed for the storage of platelets for transfusion. At present, platelets suspended in approximately 70% synthetic medium and 30% plasma can be stored for at least 5 days at very stable pH levels, generally pH 6.8-7.2. Present knowledge suggests that synthetic media should contain at least acetate, citrate, phosphate, potassium and magnesium. Future studies will probably result in the inclusion of other components to this list. Glucose for platelet metabolism will generally be supplied by carryover of plasma from the original platelet preparation. In addition, improved plastic containers for the storage of platelets will probably facilitate the introduction of new synthetic media. In six studies comparing synthetic media with plasma as the storage environment, and involving patients with intensive chemotherapy for haematological malignancies, the clinical outcome in terms of corrected count increments (CCI) generally indicated similar results. Three studies suggested significant reduction of the incidence of transfusion reactions of platelets suspended in synthetic media as compared with plasma. For future comparisons of platelet storage in either plasma or new synthetic media, additional platelet survival and recovery studies, as well as patient-transfusion studies, will be needed as in vitro data may not always reflect the clinical outcome. This will add further knowledge to data from the present few clinical studies available that compare storage of platelets in either synthetic media or plasma.