Characterization of blood components prepared from whole-blood donations after a 24-hour hold with the platelet-rich plasma method

BACKGROUND: The preparation of platelet (PLT) concentrates (PCs) from PLT-rich plasma (PRP) requires that whole blood (WB) be processed within 8 hours of collection. Increasing WB storage time to 24 hours would be logistically attractive. This study compares the in vitro quality of blood components prepared from WB stored for 8 and 24 hours at room temperature before processing with the PRP method. STUDY DESIGN AND METHODS: WB units were collected from ABO-matched blood donors. To reduce individual variations, paired donations were drawn in parallel, pooled, and split back in the collection bag. One unit was held for 6 to 8 hours and the other for 22 to 24 hours at 20 to 24 degrees C. Prestorage leukoreduced components were prepared with the PRP as intermediate product and analyzed during storage. RESULTS: RBC units prepared after an 8- or 24-hour hold were comparable in terms of hemolysis, sodium, pH, and ATP levels. RBC 2,3- diphosphoglycerate (2,3-DPG) was significantly lower in RBCs prepared from 24-hour hold donations immediately after processing but not after 20 days of storage. Residual white blood cells were approximately fivefold higher (p < 0.05) in 24-hour RBC units. For PCs, measurements for glucose, ATP, lactate, pH, extent of shape change, hypotonic shock response, and CD62p activation were similar. No differences were observed in the von Willebrand factor, factor (F)V, FVIII, and fibrinogen content of fresh-frozen plasma. CONCLUSIONS: The decrease in FVIII and RBC 2,3-DPG can be acceptable as a compromise to improve blood component logistics, but leukoreduction efficiency must be improved before considering the adoption of an overnight storage of WB before PRP processing.     

Active cooling of whole blood to room temperature improves blood component quality

BACKGROUND: Many countries use cooling plates to actively cool collected whole blood (WB) to room temperature. Until now, no paired comparison had been performed, and it was our aim to compare the effect of active versus no active cooling on the in vitro quality of WB and subsequently prepared blood components. STUDY DESIGN AND METHODS: Two units of WB were pooled and divided shortly after donation. One unit was placed under a butane‐1,4‐diol plate to obtain active cooling; the other was placed in an insulated box with other warm units to mimic worst‐case holding conditions. WB was held overnight and processed into a white blood cell (WBC)‐reduced red blood cells (RBCs), buffy coat (BC), and plasma. The BCs were further processed into platelet (PLT) concentrates. RBCs were stored for 42 days, and PLT concentrates for 8 days (n = 12 paired experiments). RESULTS: After overnight storage, ATP content of the RBCs was 4.9 ± 0.3 µmol/g Hb for actively cooled WB versus 4.5 ± 0.4 µmol/g Hb for not actively cooled WB (p < 0.001). On Day 42 of storage, RBCs prepared from this WB contained 3.1 ± 0.3 µmol ATP/g Hb with active cooling versus 2.6 ± 0.3 µmol/g Hb without (p < 0.001). Hemolysis on Day 42 was 0.35 ± 0.08% with active cooling and 0.67 ± 0.21% without (p < 0.001). No effect was observed on the in vitro quality of plasma, BC, or PLT concentrates. CONCLUSIONS: Active cooling of WB results in improved ATP levels and less hemolysis in WBC‐reduced RBCs, although the clinical implications are unclear. It has no effect on the in vitro quality of plasma or PLT concentrates.     

Characterization of blood components separated from donated whole blood after an overnight holding at room temperature with the buffy coat method

BACKGROUND: With buffy coat (BC) processing of whole blood (WB) donations, increase in WB storage time to facilitate overnight holding before the separation of blood components would be a logistically attractive development. This study undertakes a comparative in vitro characterization of blood components prepared from WB samples that were either processed within 8 hours or stored overnight at room temperature before processing by the BC method. STUDY DESIGN AND METHODS: The WB units (400 mL) collected were either processed within 8 hours (fresh blood) or stored overnight (overnight blood) at room temperature. WB units were separated into individual‐component red blood cells (RBCs), BC, and plasma. The in vitro quality of these blood components (RBCs, pooled platelet concentrates [PCs], and plasma) was analyzed during storage. RESULTS: Levels of 2,3‐diphosphoglycerate (2,3‐DPG) were found to be significantly lower immediately after processing, compared with the fresh WB samples, in RBCs that had been separated from an overnight‐hold sample. However, this difference was not apparent after 14 days of storage. In pooled PCs, measurements for glucose, lactate, PO₂, PCO₂, extent of shape change, and hypotonic shock response were similar. The platelet yield in PCs prepared from an overnight‐hold WB sample was significantly higher, while CD62P expression and annexin V binding were lower (p < 0.05). For frozen plasma (FP), no significant differences were observed for the coagulation factors (F)II, FVII, FV, F IX, FX, and FXI; fibrinogen; and von Willebrand factor content between the 8‐ and 24‐hour FP. The FVIII was the component that was most sensitive to the prolongation of production time and it only had 80% of the activity of the 8‐hour FP. CONCLUSION: These data suggest that blood components (RBCs, pooled PCs, and FP) separated from WB that has been stored overnight at room temperature by the BC method are of acceptable quality.     

Alkaline CPD and the preservation of RBC 2,3-DPG

BACKGROUND: Concentrations of 2,3-DPG decline rapidly in the first week of RBC storage because of the low pH of conventional storage solutions. Alkaline additive solutions, which can preserve RBCs for up to 11 weeks, still do not preserve 2,3-DPG because the starting pH is below 7.2. STUDY DESIGN AND METHODS: Alkaline CPD (pH=8.7) was made with trisodium citrate, dextrose, and disodium phosphate. Twelve units of whole blood were collected into heparin and pooled in groups of four units. Each pool was then aliquoted into four units; 63 mL of CPD with pH 5.7, 6.5, 7.5, or 8.7 was added to one unit of each pool, and 300 mL of the alkaline experimental additive solution-76 was added. In Study 2, 12 units were collected into alkaline CPD, pooled in groups of four, aliquoted as described, and stored in four variants of experimental additive solution-76 containing 0, 9, 18, and 27 mM of disodium phosphate. RBC ATP and 2,3-DPG concentrations, intracellular and extracellular pH and phosphate concentrations, hemolysis, and other measures of RBC metabolism and function were measured weekly. RESULTS: RBCs stored in more alkaline conditions made 2,3-DPG, but at the expense of ATP. Concentrations of 2,3-DPG decreased after 2 weeks storage, but ATP concentrations never fully recovered. Providing more phosphate both increased the duration of 2,3-DPG persistence and raised ATP concentrations in the later stages of storage. CONCLUSIONS: Maintaining both 2,3-DPG and ATP requires both high pH and high concentrations of phosphate.     

Evaluation of nonleukoreduced red blood cell transfusion units collected at delivery from the placenta

BACKGROUND: The objective of this study was to evaluate the suitability of cord blood (CB) as a source of red blood cells (RBCs) for autologous transfusion. STUDY DESIGN AND METHODS: CB was collected in 150-mL storage containers with citrate phosphate dextrose (CPD) as anticoagulant and stored in either saline, adenine, glucose, and mannitol (SAG-M; n = 18) or phosphate, adenine, glucose, guanosine, saline, and mannitol (PAGGS-M; n = 18) for 35 days at 4 degrees C. Hematologic status and hemolysis were studied. The lipopolysaccharide (LPS)-induced production of tumor necrosis factor (TNF)-alpha and transforming growth factor (TGF)-beta1 from CB monocytes was analyzed after incubation with addition of weekly sampled supernatants from the CB RBC units. Five additional units (PAGGS-M) were leukoreduced and thereafter analyzed as indicated above. RESULTS: Hemolysis increased significantly over time, in SAG-M more than in PAGGS-M. During storage in both media, the number of white blood cells (WBCs) decreased, and the LPS-induced production of TNF-alpha and TGF-beta1 decreased and increased, respectively. There were no significant changes in the LPS-induced production of TNF-alpha and TGF-beta1 in the leukoreduced CB RBC units. CONCLUSION: Hemolysis in CB RBC units increased significantly over time, and PAGGS-M appears to be superior to SAG-M as a preservation solution for CB RBC. The changes in LPS-induced TNF-alpha and TGF-beta1 production over time were probably caused by substances released from apoptotic and/or necrotic WBCs. Further studies are needed to identify both which substances are responsible for the changes in LPS-induced cytokine release and the clinical significance hereof.     

Effects of rejuvenation and frozen storage on 42-day-old AS-3 RBCs

BACKGROUND: A storage period of 42 days has been approved by the FDA for RBCs stored in NUTRICEL (AS-3). This study was undertaken to provide data to the FDA about the feasibility of salvaging AS-3 RBCs at the end of their storage period by rejuvenation and freezing, and to evaluate the effect of rejuvenation on indicators of RBC function. STUDY DESIGN AND METHODS: Healthy adults (n = 22) donated 450 mL of whole blood, and RBC components were prepared at two study sites. The components were stored at 1 to 6 degrees C for either 41 days (nonrejuvenated frozen controls, n = 6) or 42 days (rejuvenated frozen study group, n = 10; nonrejuvenated nonfrozen controls, n = 6). Rejuvenated study components and nonrejuvenated frozen controls were stored at -70 degrees C for longer than 2 weeks. Frozen units were then deglycerolized and kept for an additional 24 hours at 1 to 6 degrees C. RESULTS: 2,3-DPG, and ATP were reduced after 42-day storage to near 0 and 65 percent, respectively, of their original values. After rejuvenation and deglycerolization, the mean ATP level was 146 percent and the mean 2,3-DPG was 115 percent. The percent freeze-thaw-wash recovery was similar for rejuvenated and nonrejuvenated RBCs. Trace amounts of hypoxanthine and inosine were detected in rejuvenated units. The mean 24-hour survival (single- or double-label technique) of all components exceeded 75 percent. The t1/2 of study and control RBCs was similar. CONCLUSION: The ability of 42-day-old AS-3 RBCs to deliver oxygen after rejuvenation and freezing is not impaired. These data indicate that rejuvenated AS-3 RBCs can provide a safe and beneficial blood component immediately upon infusion.     

Storage of red blood cells with improved maintenance of 2,3-bisphosphoglycerate

BACKGROUND: During storage, red blood cells (RBCs) rapidly lose 2,3-bisphosphoglycerate (2,3-DPG) leading to an increase in the affinity for O(2) and a temporary impairment of O(2) transport. Recent clinical evaluations indicate that the quality of transfused RBCs may be more important for patient survival than previously recognized. STUDY DESIGN AND METHODS: Glucose-free additive solutions (ASs) were prepared with sodium citrate, sodium gluconate, adenine, mannitol, and phosphates at high pH, a solution that can be heat-sterilized. CP2D was used as an anticoagulant. Additional CP2D was added to the AS to supply glucose. RBCs were stored at 4 degrees C and assayed periodically for intracellular pH (pHi), extracellular pH, glucose, lactate, phosphate, ATP, 2,3-DPG, hemolysis, and morphology. RESULTS: Storage in 175 mL of the chloride-free, hypotonic medium at a hematocrit (Hct) level of 59 to 60 percent resulted in an elevated pHi and the maintenance of 2,3-DPG at or above the initial value for 2 weeks without loss of ATP. The addition of 400 mL of storage solution followed by centrifugation and removal of 300 mL of excess solution to a Hct level of 60 to 66 percent further reduced the chloride concentration, resulting in the maintenance of 2,3-DPG for 4 weeks. Hemolysis was at 0.1 percent at 6 weeks. CONCLUSION: Improvements in the maintenance of 2,3-DPG were achieved with 175 mL of a chloride-free storage solution with familiar additives at nontoxic concentrations to increase pHi. Adding, instead, 400 mL of storage solution followed by the removal of 300 mL reduced the chloride concentration, increasing the pHi and extending the maintenance of 2,3-DPG to 4 weeks.     

Preparation and storage of white blood cell-reduced split apheresis platelet concentrates for pediatric use

BACKGROUND: White blood cell (WBC) reduction and bacterial screening induce unacceptable product loss when platelet (PLT) concentrates (PCs) for pediatric transfusion are prepared from whole blood. The aim was to investigate PCs, WBC reduced and bacterially screened, from single-donor apheresis procedures, divided in 3 or 4 pediatric units and stored up to 5 days. STUDY DESIGN AND METHODS: PCs were collected with an apheresis machine and WBC reduced by in-process filtration. The PCs were sampled for bacterial screening and subsequently divided in 70-mL products. Initially, storage characteristics of split units in 400-mL polyvinylchloride (PVC) bags with 17 split PCs originating from five apheresis donations were studied. When a 600-mL container made of the more gas-permeable polyolefin became available, a paired comparison was performed with 9 split PCs from nine donations and with a higher-yield PLT collection procedure. RESULTS: Split PCs contained 69 x 10(9) +/- 14 x 10(9) PLTs in 69 +/- 1 mL of plasma, and storage in the PVC containers gave a pH value of 6.86 +/- 0.10 on Day 6 (mean +/- SD, n = 17). When comparing the containers, the PVC bag contained 98 x 10(9) +/- 15 x 10(9) PLTs in 72 +/- 4 mL versus 102 x 10(9) +/- 18 x 10(9) PLTs in 74 +/- 8 mL for the polyolefin bag (n = 9, not significant). This gave pH values on Day 6 of 6.12 +/- 0.50 in the PVC container, whereas pH remained acceptable in the polyolefin container: 6.85 +/- 0.10 on Day 6 (p < 0.01). CONCLUSION: PCs for pediatric use from split single-donor apheresis concentrates, WBC reduced and bacterially screened, can be stored for up to 5 days in a 600-mL polyolefin container with maintenance of good in vitro storage variables.     

Immune response to autologous transfusion in healthy volunteers: WB versus packed RBCs and FFP

BACKGROUND: Storage of blood as packed RBCs and FFP is standard practice in allogeneic transfusion. Separation into components has been proposed for autologous transfusion, as well, but beneficial effects have not yet been shown. STUDY DESIGN AND METHODS: Twenty-four healthy male volunteers were randomly assigned to receive 1 unit of either autologous RBCs and FFP (RCP group) or WB (WB group) after 49 or 35 days of storage, respectively. The immune response was analyzed by ELISA for IL-6, C3a, terminal complement complex SC5b-9, TNF-alpha, and neopterin. Differential WBC counts and the phagocytosis of neutrophils and monocytes were measured by flow cytometry. RESULTS: Cell counts of monocytes (0.85 x 10(3) ng/microL) [corrected] and neutrophils (6.9 x 10(3) ng/microL) [corrected] increased 30 minutes after WB transfusion and then returned to close to the baseline values seen in the RCP group (0.47 and 2.9 x 10(3) ng/microL [corrected], respectively) throughout the monitored period (p<0.05). C3a (169 vs. 116 ng/microL) [corrected] and IL-6 (29 vs. 6 pg/mL) reached higher plasma concentrations in the WB group (n = 11) than in the RCP group (n = 10). Phagocytosis of opsonized Escherichia coli was increased in neutrophils and monocytes and lasted up to 7 days after the transfusion of whole blood. CONCLUSION: Autologous WB induces a modest immunomodulation, but this effect is not observed upon transfusion of autologous blood components.     

Effects of different concentrations of anticoagulant on the in vitro characteristics of autologous whole blood

BACKGROUND: Routinely, 450 mL of blood is collected into 63 mL of CPDA-1, for a final anticoagulant:blood ratio of approximately 1:7 in a whole-blood autologous unit. If less than 300 mL of blood is to be collected, the AABB standards suggest that there should be a proportionate decrease in anticoagulant. Data from an autologous blood program showed a range in volume from 92 mL to 667 mL per bag, which reflects an anticoagulant:blood ratio of 2:1 to 1:10. STUDY DESIGN AND METHODS: To determine the effects of these ratios on the in vitro function of RBCs at various anticoagulant ratios, blood was collected into different amounts of anticoagulant, and various measurements were made during storage. RESULTS: The number of RBCs and the MCV remained constant over time, regardless of the anticoagulant dilution used. Plasma free Hb increased with time with all dilutions. At a 1:2 ratio, it rose from 734 mg per L on Day 1 to 1805 mg per L on Day 35, and at 1:8, it was 355 mg per L for Day 1 and 854 mg per L on Day 35. Plasma sodium decreased and the potassium increased over time with all dilutions. From Day 1 to Day 35, there was a nine-fold increase in potassium at both the 1:2 and 1:8 dilutions (2.4 to 22.9 mmol/L, 3.2 to 29.6 mmol/L, respectively). The LDH increased over time and the pH decreased in all of the dilutions. Osmotic fragility remained constant at the 1:8 dilution but decreased at all of the other dilutions with storage, with 44-percent fragility on Day 35 at the 1:2 ratio. The WBC and platelet counts decreased consistently over time. Overall, 1 percent of the autologous units were below the cutoff volume of 300 mL at which an adjustment of the anticoagulant volume is required. CONCLUSION: Plasma Hb and plasma potassium concentrations are considerably higher in low-volume units, which indicates that deviation from standard collection procedures is deleterious to RBCs.     

Autologous red cells derived from cord blood: collection, preparation, storage and quality controls with optimal additive storage medium (Sag-mannitol)

To investigate whether packed red cells (PRCs) prepared from autologous cord blood-packed red cells (AC-PRCs) could be used as an alternative for homologous-packed red cells (H-PRCs), we developed a system to collect and prepare AC-PRCs and determined standard storage parameters during 35 days of storage in extended storage medium (Sag-mannitol). We collected and fractionated cord blood from 390 newborns. The amount and quality of the AC-PRCs were analysed. The bacterial contamination rate was 1.84%. Twelve AC-PRCs were stored for 35 days, and standard laboratory parameters were measured at day 1 and day 35. The initial laboratory parameters of the AC-PRCs were similar to the parameters of the H-PRCs. After 35 days, the AC-PRCs displayed an increased haemolysis rate compared to H-PRCs (1.1 versus 0.2%) and also a significant decreased adenosine triphosphate value (1.2 versus 2.3 micromol L(-1)). Haemoglobin, haematocrit and pH were comparable in both groups. AC-PRCs meet the quality criteria for H-PRCs after 35 days. Utilizing a closed collection system for cord blood and an extended storage medium will increase safety and quality and facilitate the routine transfusion of autologous red cells derived from cord blood.     

In vivo viability of stored red blood cells derived from riboflavin plus ultraviolet light-treated whole blood

BACKGROUND: A novel system using ultraviolet (UV) light and riboflavin (Mirasol System, CaridianBCT Biotechnologies) to fragment nucleic acids has been developed to treat whole blood (WB), aiming at the reduction of potential pathogen load and white blood cell inactivation. We evaluated stored red blood cell (RBC) metabolic status and viability, in vitro and in vivo, of riboflavin/UV light–treated WB (IMPROVE study). STUDY DESIGN AND METHODS: The study compared recovery and survival of RBCs obtained from nonleukoreduced WB treated using three different UV light energies (22, 33, or 44 J/mL_RBC). After treatment, WB from 12 subjects was separated into components and tested at the beginning and end of component storage. After 42 days of storage, an aliquot of RBCs was radiolabeled and autologously reinfused into subjects for analysis of 24‐hour recovery and survival of RBCs. RESULTS: Eleven subjects completed the in vivo study. No device‐related adverse events were observed. By Day 42 of storage, a significant change in the concentrations of sodium and potassium was observed. Five subjects had a 24‐hour RBC recovery of 75% or more with no significant differences among the energy groups. RBC t_1/2 was 24 ± 9 days for the combined three groups. Significant correlations between 24‐hour RBC recovery and survival, hemolysis, adenosine triphosphate (ATP), and CO₂ levels were observed. CONCLUSIONS: This study shows that key RBC quality variables, hemolysis, and ATP concentration may be predictive of their 24‐hour recovery and t_1/2 survival. These variables will now be used to assess modifications to the system including storage duration, storage temperature, and appropriate energy dose for treatment.     

Level of platelet-derived cytokines in leukoreduced red blood cells is influenced by the processing method and type of leukoreduction filter

BACKGROUND: In contrast to the well-documented effect of white blood cells on the quality of red blood cells (RBCs), the effect of platelets (PLTs) has received little consideration. In this study, the PLT content and level of PLT-derived cytokines in RBCs prepared using different types of leukoreduction methods were investigated.
STUDY DESIGN AND METHODS: Buffy coat–poor RBCs and five types of leukofiltered (LF) RBCs, including RBCs prepared with a whole blood (WB) PLT-saving filter, were prepared and stored according to standard blood bank conditions. PLT content was measured on Day 1, and levels of PLT-derived cytokines were measured by enzyme-linked immunosorbent assay at nominated timepoints during 42 days of storage.
RESULTS: The PLT content of leukoreduced RBCs varied widely depending on the processing method and/or leukoreduction filter used, with some types of RBCs containing very low PLT counts while other units contained PLT counts comparable to those of unprocessed WB. The PLT content of RBCs directly influenced the concentration and accumulation of PLT-derived cytokines. Several PLT-derived factors exhibited significant accumulation throughout 42 days of storage. RBCs with high PLT content exhibited concentrations of RANTES (CCL5) and soluble CD40 ligand equivalent to those previously reported to show significant biologic and clinical effects.
CONCLUSION: The PLT content and levels of PLT-derived cytokines in leukoreduced RBCs are influenced by the processing method and types of leukoreduction filters used. It may be inappropriate to consider LF-RBCs prepared with different types of leukoreduction filters as equivalent products based on their differing levels of PLT factors.     

Prolonged maintenance of 2,3-diphosphoglycerate acid and adenosine triphosphate in red blood cells during storage

BACKGROUND: Current additive solutions (ASs) for red cells (RBCs) do not maintain a constant level of critical metabolites such as adenosine triphosphate (ATP) and 2,3-diphosphoglycerate acid (2,3-DPG) during cold storage. From the literature it is known that the intracellular pH is an important determinant of RBC metabolism. Therefore, a new, alkaline, AS was developed with the aim to allow cold storage of RBCs with stable product characteristics. STUDY DESIGN AND METHODS: Whole blood-derived RBCs (leukoreduced) were resuspended in experimental medium phosphate-adenine-guanosine-glucose-gluconate-mannitol (PAGGG-M; pH 8.2) with and without washing in the same medium. During cold storage several in vitro variables, such as intracellular pH, 2,3-DPG, ATP, and hemolysis, were analyzed. RESULTS: During cold storage, RBCs resuspended in PAGGG-M showed a constant ATP level (approx. 6 mumol/g Hb) and a very limited hemolysis (<0.2%). The 2,3-DPG content showed an increase until Day 21 (150% of initial level), followed by a slow decrease, with at Day 35 still 100 percent of the initial level. RBCs washed in PAGGG-M even showed a continuous increase of 2,3-DPG during 35 days, with a maximum level of 200 percent of the initial value. The effect of PAGGG-M appears to be related to long-lasting effects of the initial intracellular pH shortly after production. CONCLUSION: Resuspension of RBCs in our alkaline medium PAGGG-M resulted in a RBC unit of high quality during storage for up to at least 35 days, with 2,3-DPG levels of higher than 10 mumol per g Hb, hemolysis of less than 0.2 percent, and ATP levels of higher than 5 mumol per g Hb.     

The influence of maintaining the correct whole blood-to-anticoagulant ratio during donation on the quality of leukoreduced whole blood

BACKGROUND: It is unclear whether maintaining the correct whole blood‐to‐anticoagulant (WB : AC) ratio during collection can improve the quality of red blood cell (RBC)‐containing blood products to a clinically relevant degree. STUDY DESIGN AND METHODS: A total of 2 × 20 CPDA‐1 leukoreduced whole blood (WB) units suspended in CPDA‐1 were investigated. In one group, AC was continuously added to the donated blood, maintaining the correct WB : AC ratio during collection, using a new drawing device (MacoPharma ABC). In the other group, WB units were produced conventionally. Adenosine triphosphate (ATP), 2,3‐diphosphoglycerate, free hemoglobin (Hb), potassium, glucose, lactate, pH, and variables of coagulation were determined on Days 1, 7, 21, 35, 42, and 49 of storage. Variables of RBC deformability and aggregability were determined using a laser‐assisted optical rotational cell analyzer. RESULTS: The ABC and conventional group showed comparable unit volumes of 525 (SD, 5.3) mL versus 524 (SD, 10.2) mL and Hb content of 65.9 (SD, 5.1) g/unit versus 67.5 (SD, 7.8) g/unit, but higher variation after conventional blood drawing (p = 0.006 and p = 0.07, respectively) was observed. During storage, none of the measured quality variables were significantly different between the groups. Mean (SD) ATP was 2.33 (0.41) µmol/g Hb versus 2.24 (0.39) µmol/g Hb after 42‐day storage. Deformability was not different (p = 0.44), whereas the extent of the aggregability was higher in the conventional group (p = 0.04). CONCLUSION: The ABC device provided a better standardized blood product but did not improve RBC storage variables or plasma quality. It slightly reduced RBC aggregability during storage. Excess AC at the beginning of a donation appears not to significantly affect RBC storage in conventional blood drawing.     

The 24-hour posttransfusion survival of baboon red blood cells preserved in citrate phosphate dextrose/ ADSOL (CPD/AS-1) for 49 days.

The purpose of this study was to evaluate the baboon as an animal model for evaluating red blood cell (RBC) preservation by comparing the 24-h posttransfusion survival of baboon RBCs preserved in citrate phosphate dextrose/ADSOL (CPD/AS-1) solution at 4 degrees C for 49 days to that of human RBCs preserved under similar conditions. CPD/AS-1 originally was approved by the Food and Drug Administration for 49-day storage of RBCs, but this period subsequently was reduced to 42 days. Adult male baboons (Papio anubis and P. cynocephalus) were autotransfused with RBCs that had been harvested using CPD and that had been resuspended and stored in AS-1 solution at 4 degrees C for as long as 49 days. The 24-h posttransfusion survival was measured using the 51Cr/125I-albumin method. The 24-h posttransfusion survival (mean +/- standard deviation) was 74% +/- 7% for seven units of CPD/AS-1-treated RBCs stored for 35 days, 65% +/- 15% for 12 units stored for 42 days, and 43% +/- 16% for seven units stored for 49 days. The mean 24-h posttransfusion survival rate for autologous baboon RBCs stored in CPD/AS-1 at 4 degrees C for 35 days (74%) was similar to that for autologous human RBCs stored in a similar manner. Further storage for 42 and 49 days resulted in lower values for baboon RBCs compared with human RBCs.     

Quality and stability of red cells derived from gravity-separated placental blood with a hollow-fiber system

BACKGROUND: Several studies show that donor red blood cells (RBCs) can be processed by gravity separation with a hollow-fiber filtration system. This study investigated whether fetal blood could be filtered in the same way. STUDY DESIGN AND METHODS: Twelve newborns born after healthy pregnancies were included in the study. Placental blood was sampled with standard procedures. The sampled blood was separated with a specially designed hollow-fiber filtration system (Sangofer neonatal, Heim Group). The RBC bag contained 10 mL of saline, adenine, glucose-mannitol (SAG-M) for stabilization. After processing, the resulting RBC volume was estimated. Quality variables (blood counts, hemolysis rate) were measured before and after 35 days of storage at +4 degrees C. RESULTS: The 12 processed RBC units had a mean volume of 62.3 +/- 13.5 mL and a mean hematocrit level of 0.56 +/- 0.06. No white blood cell contamination could be detected in any of the RBC units tested. After 35 days of storage, the hemolysis was 0.1 +/- 0.07 and the amount of free hemoglobin was 0.28 +/- 0.017 mmol per L. CONCLUSIONS: This study shows that it is possible to process placental blood to RBCs by gravity separation with a hollow-fiber system. The quality of the RBCs thus processed was suitable for 35 days storage. The use of placental blood in the treatment of children with anemia (e.g., malaria) in the underresourced world is widely discussed. Because the separation device used here needs no additional equipment or electrical devices, it is considered to be an ideal method for use in these countries.     

Cord blood as a source of autologous RBCs for transfusion to preterm infants

BACKGROUND: This prospective study was conducted to gain experience as to whether it is technically possible to produce autologous RBCs in additive solution from cord blood (CB), to optimize the blood supply for preterm infants. STUDY DESIGN AND METHODS: CB was collected from 47 infants with a mean (+/- SD) birth weight of 1717 (+/- 699) g. Whenever possible, RBC components were prepared by standard centrifugation using a six-bag system. All samples were put in sterility testing quarantine for 5 days, and a maximum storage of 14 days from collection to transfusion was specified. The babies were given either the autologous RBCs or standard allogeneic RBC concentrates, if autologous blood was not available. RESULTS: In 81 percent of the samples, autologous RBC components could be processed (vol, 7-87 mL; Hct, 31-82%). But within the group of extremely low birth weight infants (body weight <1000 g), a mean CB net volume of only 37 mL was collected, and the RBC preparation was successful only in exceptional cases. Three CB samples (8.6%) tested positive in sterility testing. Of the 47 infants, 21 were treated with a total of 62 allogeneic and 4 autologous RBC transfusions. Most infants with a body weight over 1400 g did not need any RBC transfusion. CONCLUSION: The preparation of autologous RBCs from the CB of preterm infants is technically possible in principle. However, major concerns must be raised as to whether such preparations are of benefit in ensuring safe care of neonates with blood components, with respect to the high rate of bacterial contamination and the limited availability in babies with low birth weight.     

Rejuvenation capacity of red blood cells in additive solutions over long-term storage

BACKGROUND: Red blood cells (RBCs) are Food and Drug Administration (FDA)‐approved for 42‐day storage with the use of additive solutions (ASs). However, adenosine triphosphate (ATP) and 2,3‐diphosphoglycerate (2,3‐DPG) levels in the RBCs decline over this time. These constituents may be restored by treatment with rejuvenation (REJ) solutions. This study was done to assess the response capability of RBCs from 30 to 120 days of storage in three FDA‐licensed RBC storage solutions after incubation with a rejuvenating solution of pyruvate, inosine, phosphate, and adenine. STUDY DESIGN AND METHODS: Three units each of RBCs in approved AS (AS‐1 [Adsol, Fenwal, Inc.], AS‐3 [Nutricel, Medsep Corp.], and AS‐5 [Optisol, Terumo Corp.]) were stored under standard conditions at 1 to 6°C for up to 120 days. Aliquots (4 mL) on Days 30, 42, 60, 80, 100, and 120 (±2 days) were REJ by incubating with Rejuvesol (Encyte Corp.). Control untreated and REJ aliquots were extracted using perchloric acid and stored at ?80°C until assayed for 2,3‐DPG and ATP. RESULTS: RBCs responded to REJ by increasing DPG and ATP contents. The response declined linearly at 0.070 ± 0.008 µmol DPG/g hemoglobin (Hb)/day and 0.035 ± 0.004 µmol ATP/g Hb/day with no differences between ASs. CONCLUSION: We conclude that Rejuvesol is able to restore ATP and 2,3‐DPG levels in RBCs stored up to 120 days in AS. The response diminishes as storage time increases. This rejuvenation (REJ) capability does not seem useful for routine assessment of RBC anabolic capacity in research programs, but may be useful to the investigator when studying unique and novel treatment methods.     

Manufacture of red cells in additive solution from whole blood refrigerated for 5 days or remanufactured from red cells stored in plasma

Background and Objectives: To investigate methods for the production of red cell concentrates (RCC) in saline, adenine, glucose and mannitol (SAG‐M), from whole blood or red cells stored in plasma for 5 or 6 days and to provide evidence that exchange transfusion RCC in citrate phosphate dextrose (CPD) plasma or citrate, phosphate, dextrose, adenine (CPDA‐1) plasma are of comparable quality. Methods and Materials: Ten RCC in SAG‐M were produced following the remanufacture of red cells in CPD plasma on day 5/6 or after 5 days hold as leucodepleted CPD whole blood. In addition, 10 RCC in CPD plasma and 9 in CPDA‐1 plasma were stored without further processing. Units were assessed for red cell parameters including haemolysis, adenosine triphosphate (ATP), 2,3‐diphosphoglycerate (2,3‐DPG) and extracellular potassium. Results: Units in SAG‐M produced by remanufacture of RCC in plasma or by delayed manufacture of whole blood had comparable levels of haemolysis, ATP and 2,3‐DPG. Furthermore, these units underwent biochemical changes similar to reference SAG‐M units, with the exception of haemolysis which was greater at the end of shelf life and supernatant potassium which was lower following remanufacture. As expected, the decline in ATP was greater in red cells stored in CPD plasma compared with CPDA‐1 plasma. In general, units in CPD plasma were of similar quality at day 28 compared to those in CPDA‐1 plasma at day 35. Conclusions: RCC produced following the remanufacture of RCC in plasma or the delayed manufacture of whole blood are of acceptable in vitro quality and should be assigned the same shelf life as standard RCC in SAG‐M.