The quality of fresh-frozen plasma produced from whole blood stored at 4 degrees C overnight

BACKGROUND: The aim of this study was to assess whether the quality of FFP produced from whole blood stored at 4 degrees C overnight is adequate for its intended purpose. STUDY DESIGN AND METHODS: Fresh-frozen plasma (FFP) separated from whole blood (n = 60) leukodepleted (LD) after storage at 4 degrees C overnight (18-24 hr from donation, Day 1 FFP) was compared with that LD within 8 hours of donation (Day 0 FFP, the current standard method). RESULTS: In more than 95 percent of Day 1 FFP units, levels of factor (F) II, FV, FVII, FVIII, F IX, FX, FXI, and FXII were greater than 0.50 U per mL except for von Willebrand factor (VWF) antigen and FVIII, where 92 and 87 percent of units, respectively, contained greater than 0.50 IU per mL. Compared with historical data on FFP stored for 8 hours, fibrinogen, FV, FVIII, and FXI were reduced by 12, 15, 23, and 7 percent, respectively, but other factors were not significantly reduced. Levels of VWF-cleaving protease activity were not different between FFP prepared from paired units of blood (n = 3) held for 8 or 24 hours, but were below the reference range in an additional 2 of 6 units held for 24 hours. The activities of protein S, protein C, antithrombin III, and alpha(2)-antiplasmin were reduced by less than 10 percent in Day 1 FFP (n = 20), but with final levels above the lower limit of the normal range in greater than 95 percent of units. Activated FXII antigen was not significantly raised in plasma stored for 18 to 24 hours, but levels of prothrombin fragment 1 + 2 were slightly increased (0.88 ng/mL, 18-24 hr; 0.65 ng/mL, < 8 hr). CONCLUSION: These data suggest that there is good retention of relevant coagulation factor activity in plasma produced from whole blood stored at 4 degrees C for 18 to 24 hours and that this would be an acceptable product for most patients requiring FFP.     

Effect of an 8-hour holding period on in vivo and in vitro properties of red cells and factor VIII content of plasma after collection in a red cell additive system

Extension of the holding time for whole blood units from 6 to 8 hours at ambient temperature should provide enhanced flexibility in the preparation of platelet concentrates (PCs). A paired study was conducted to evaluate the characteristics of stored red cells (RBCs) and plasma prepared from whole blood collected into a red cell additive system (CPD-ADSOL) after an extended holding time. An individual donated a unit of whole blood on two occasions; 1 unit was held for 6 hours before processing and the other for 8 hours. Autologous RBC 24-hour survival levels after 42 days of storage were comparable. Laboratory A, using a 99mTc-51Cr technique, found mean survival levels of 79 percent (6-hour hold) and 78 percent (8-hour hold) (n = 8). Analysis by the single-label procedure found the mean levels to be 82 and 81 percent. Laboratory B, using an albumin 125I-51Cr technique, found mean survival levels of 74 and 72 percent (n = 10). Mean hemolysis and ATP levels were found to be comparable after 42 days of storage following 6- and 8-hour holding periods. 2,3 DPG levels were reduced to a greater degree during the longer hold. The factor VIII levels in plasma frozen for at least a month after 6- and 8-hour holding periods were comparable; thawed plasma contained mean levels of 0.77 and 0.76 units per mL (n = 21). These studies indicate that components prepared by using a CPD-ADSOL system after holding periods of 6 and 8 hours have comparable properties.     

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.     

Plasma factor VIII, variables affecting stability under standard blood bank conditions and correlation with recovery in concentrates

Some of the variables affecting factor VIII during storage of plasma at 4 C prior to preparing factor VIII concentrates and cryoprecipitates were investigated. No significant difference in factor VIII levels could be demonstrated between whole blood, platelet-rich plasma or plasma depleted of platelets when stored at 4 C. However, in frozen plasma platelets were shown to have a deleterious effect on factor VIII. No significant difference in factor VIII levels was found in whole blood or plasma stored at 4 C for 6 hours as compared with blood or plasma stored for 18 hours (p = greater than 0.1). There was, however, a highly significant difference (p = less than 0.001) between plasma and whole blood stored at 4 C for 4 hours as compared with storage for 18 hours. There was a significant correlation (correlation coefficient 0.714) between factor VIII levels in the starting material and the factor VIII recovered in cryoprecipitates, but in a small pilot study no correlation between factor VIII levels in the starting material and that recovered in freeze-dried concentrates, could be found. There was no significant difference in factor VIII levels between the Group O and Group A donations used in this study (p = greater than 0.1), and no correlation was found between the drop in pH during storage and the factor VIII decay pattern.     

Effect of 24-hour whole-blood storageon plasma clotting factors

BACKGROUND: The current requirements for the preparation of fresh-frozen plasma within 8 hours of whole-blood collection were designed to maintain clotting factor activities. These requirements, however, limit the production of fresh-frozen plasma in a large blood center. There are few data on the effect of the extension of CPD whole-blood storage to 24 hours on clotting factor activity. STUDY DESIGN AND METHODS: A 500-mL unit of whole blood was collected from 10 volunteer donors. At 1 hour after collection, a plasma sample was separated by centrifugation, and each unit was equally divided into 2 half-units, with 1 half-unit stored at 4 degrees C (range, 1-6 degrees C) and 1 half-unit stored at 22 degrees C (range, 20-24 degrees C) for 8 hours after collection. Each half-unit was then placed at 4 degrees C for further storage for 16 hours. At 8 and 24 hours after collection, plasma samples were separated from each half-unit. All plasma samples were frozen at -18 degrees C. Factors V, VII, VIII, and X; fibrinogen; antithrombin III; protein C; and protein S were measured. RESULTS: No significant changes were noted in factors V, VII, and X; fibrinogen; antithrombin III; protein C; and protein S over the 24-hour storage period. Factor VIII in both half-units was significantly reduced, by 13 percent, from the baseline sample as compared to the level in the 8-hour storage sample (p<0.05). Factor VIII was further reduced by 15 to 20 percent after the 24-hour storage period (p<0.05). CONCLUSION: The coagulation factor activity for all factors measured, with the exception of factor VIII, showed no significant change over the 24-hour storage period. Factor VIII was significantly decreased by 13 percent in 8-hour storage and by an additional 15 to 20 percent in 24-hour storage. For clinical situations not requiring the replacement of factor VIII only, 24-hour frozen plasma has properties comparable to those of fresh-frozen plasma.     

Bacteria levels in components prepared from deliberately inoculated whole blood held for 8 or 24 hours at 20 to 24 degrees C

BACKGROUND : An increase from 8 to 24 hours in the time that units of whole blood can be held at room temperature after phlebotomy would give blood centers more flexibility in component manufacturing and might allow receipt of many infectious disease test results prior to component preparation. However, the potential for bacterial growth during prolonged holding periods requires further study. STUDY DESIGN AND METHODS : In the Phase I study, 2-unit pools of ABO-identical whole blood were deliberately inoculated on Day 0 with Staphylococcus aureus or Pseudomonas fluorescens. They were then divided in half and stored at 20 to 24 degrees C. Red cells (RBCs) with additive solution, platelet concentrates (PCs), and frozen plasma were prepared after 8 and 24 hours. Bacteria levels in PCs and RBCs were monitored on Day 1; bacteria levels were measured in plasma after thawing. In the Phase II study, the same basic design as in Phase I was used, except that 10 bacterial species were studied, lower inocula were used, and RBCs prepared after a 24-hour room-temperature whole-blood hold were white cell-reduced by filtration. Bacterial growth was monitored during 42- day storage of RBCs (1 – 6 degrees C) and 5-day storage of PCs (20 – 24 degrees C) and after thawing of frozen plasma. RESULTS : For Phase I, significantly higher bacteria levels were observed in RBCs prepared after a prolonged hold (p < 0.05); higher levels were not observed in PCs and thawed plasma units. In Phase II, prior to white cell reduction by filtration, 8 of 10 organisms had significantly higher levels in RBCs prepared after a 24-hour hold than in RBCs prepared after an 8- hour hold, when both were examined on Day 1 (p < 0.05). For seven of eight organisms examined on Days 1, 21, and 42, filtration (white cell reduction) reduced the bacteria in RBCs prepared from 24-hour whole blood units to those levels found in unfiltered RBCs prepared from whole blood units held at 8 hours. A prolongation of the holding time from 8 to 24 hours resulted in significantly lower bacteria levels (p < 0.05) in PCs early in storage (Days 1, 1 – 2, or 1 – 3) for seven organisms, with no significant difference for two organisms, and a small but significant increase for one organism (Day 3, p < 0.05). There was no difference in bacteria or endotoxin levels in thawed units of plasma prepared from whole blood after 8- or 24-hour holding times. CONCLUSION : The levels of bacteria present in components after deliberately inoculated whole blood units are held for 8 and 24 hours depended on the organisms tested, the whole-blood holding period, and the blood component assayed; for RBCs, they also depended on whether WBC reduction by filtration was performed.     

Posttransfusion survival (24-hour) and hemolysis of previously frozen, deglycerolized RBCs after storage at 4 degrees C for up to 14 days in sodium chloride alone or sodium chloride supplemented with additive solutions

BACKGROUND: Previously frozen human RBCs currently are glycerolized and deglycerolized by the use of open systems that limit storage of the deglycerolized RBCs at 4 degrees C to only 24 hours. STUDY DESIGN AND METHODS: Healthy male volunteers who met AABB requirements for blood donors (n = 38) were studied. A volume of 450 mL of blood was collected into CPDA-1. The RBC concentrates were stored at 4 degrees C for 3 to 6 days before being frozen with 40-percent (wt/vol) glycerol and stored at -80 degrees C. The RBCs were deglycerolized, resuspended in 0.9-percent sodium chloride and 0.2-percent glucose (SG) solution or SG solution supplemented with AS-1, AS-3, or AS-5, and stored in the resuspension medium at 4 degrees C for 14 days. RESULTS: The mean +/- SD freeze-thaw-wash process recovery was 90.0 +/- 4.0 percent for all 38 units. The mean 24-hour posttransfusion survival value was 79 percent for deglycerolized RBC stored at 4 degrees C for 7 days in SG alone, SG plus AS-3, or SG plus AS-5. Deglycerolized RBC that were stored at 4 C for 14 days in SG supplemented with AS-1, AS-3, or AS-5 had a mean 24-hour posttransfusion survival of 74 percent. After 7 days of storage of deglycerolized RBCs in SG alone, the mean hemolysis was 3. 7 percent. After 14 days of storage of deglycerolized RBCs in SG supplemented with AS-1, AS-3, or AS-5, the mean hemolysis was 2.5 percent. CONCLUSIONS: The levels of hemolysis did not correlate with the 24-hour posttransfusion survival values.     

Postfiltration factor VIII and fibrinogen levels in cryoprecipitate stored at room temperature and at 1 to 6 degrees C

The 13th edition of the standards of the American Association of Blood Banks specified storage at 1 to 6 degrees C for cryoprecipitated anti-hemophilic factor (Cryo) administered up to 6 hours after thawing if the Cryo is used for factor VIII (FVIII) content (Standard J4.210). Previous editions specified room-temperature (RT) storage for up to 6 hours. Currently, the temperature specification has been deleted. There are few data addressing the optimal storage temperature and maximum storage time for FVIII and fibrinogen in thawed Cryo. Thirty bags of Cryo were assayed for FVIII and fibrinogen. Each bag was divided into two aliquots; one was stored at RT and the other at 1 to 6 degrees C. Assays were performed immediately after thawing (Base) and 6 and 24 hours after thawing, respectively. All samples were filtered through 200-mu blood component infusion sets before assay. Three hundred analyses were performed, 150 each for FVIII and fibrinogen by conventional clotting technique. Data were analyzed by using a paired t test. Cryo stored at 1 to 6 degrees C for 6 and 24 hours showed an FVIII loss of 35 percent (p less than 0.0001) and 63 percent (p less than 0.0001), respectively. Cryo stored at RT for 6 and 24 hours had an FVIII loss of 8 percent (p greater than 0.05) and 20 percent (p less than 0.0001). Cryo stored at 1 to 6 degrees C for 6 and 24 hours had a fibrinogen loss of 20 percent (p less than 0.0001) and 43 percent (p less than 0.0001). Cryo stored at RT for 6 hours had no fibrinogen loss and a 2 percent loss at 24 hours (p greater than 0.05). These preliminary data show a significant loss of FVIII and fibrinogen activity in Cryo stored at 1 to 6 degrees C and filtered before assay. The FVIII and fibrinogen activity at RT is clearly maintained up to 6 hours after thawing.     

Methods of assessing factor VIII content of stored fresh frozen plasma intended for preparation of factor VIII concentrates

Minimally destructive methods were sought to assess the factor VIII content of fresh frozen plasma intended for large-scale fractionation and stored in five-liter polyethylene packs after pooling approximately 23 plasma donations. Although factor VII distribution in the frozen pack was not perfectly homogenous, a central "core" through the frozen pack gave a representative sampel of the entire contents of the pack. Coring was compared with other methods of pack sampling before large- scale cryoprecipitation. The survival of factor VIII was studied in three grades of stored plasma which had been separated and frozen within 4 hours, 8 hours, and 18 hours of blood donation. Plasma frozen after overnight storage of blood is a satisfacory starting material for the preparation of factor VIII and factor IX concentrates.     

Survival and function of baboon RBCs released from clotted blood and washed before autologous transfusion

BACKGROUND: One alternative to an allogeneic transfusion is the salvaging of the patient's own shed blood. In this study, baboon blood was allowed to clot and the RBCs that were released from the clotted blood lysed with and without urokinase were washed before autologous transfusion. STUDY DESIGN AND METHODS: Forty-four studies were done in 13 baboons (Papio cynocephalus or Papio anubis) over a 3-year period. In 24 studies, a 50-mL volume of blood was collected without an anticoagulant and stored at 22 degrees C for as long as 72 hours before washing and autologous transfusion. In 20 other studies, a 50-mL volume of blood was collected without an anticoagulant and allowed to clot for 30 to 60 minutes. Urokinase, ranging from 2,500 to 10,000 units per mL, was added, and the blood was stored at 22 degrees C for 24 hours before washing and autologous transfusion. RESULTS: RBCs that were stored at 22 degrees C without urokinase for 24 hours exhibited an in vitro recovery value of 45 percent, a (51)Cr 24-hour posttransfusion survival of 86 percent, and an index of therapeutic effectiveness of 39 percent. The (51)Cr T(50) value was normal at 14 days, and RBC oxygen-transport function was slightly reduced. RBCs that were stored at 22 degrees C for 24 hours with 10,000 units per mL of urokinase exhibited an in vitro recovery value of 89 percent, a (51)Cr 24-hour posttransfusion survival value of 86 percent, and an index of therapeutic effectiveness of 76 percent. The (51)Cr T(50) value was normal at 14 days, and the RBC oxygen-transport function was only slightly reduced. CONCLUSION: Autologous baboon RBCs isolated from clotted blood treated or not treated with urokinase and washed before transfusion have excellent survival and normal or only slightly reduced oxygen-transport function.     

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.     

Platelet viability following storage for 5 days. Influence of holding whole blood for 8 hours at 20 to 24 degrees C before concentrate preparation

Regional blood centers frequently need to hold units of whole blood at 20 to 24 degrees C for several hours after phlebotomy so that sufficient platelet concentrates can be prepared to meet the increasing need. We have evaluated the in vivo viability and in vitro properties of platelets that were prepared from whole blood drawn into citrate- phosphate-dextrose-adenine (CPDA-1) either immediately after phlebotomy or after an 8-hour hold at 20 to 24 degrees C. Platelet concentrates were stored for 5 days at 20 to 24 degrees C in polyolefin containers (PL 732, Fenwal) with end-over-end tumbler agitation. The autologous in vivo recovery (mean +/− SD) and one-half disappearance of 51Cr-labeled platelets prepared immediately after phlebotomy were 44.4 +/− 9.4 percent and 4.0 +/− 0.5 days, respectively. Platelets prepared after the delay of 8 hours showed a recovery of 44.5 +/− 8.4 percent and a one-half disappearance of 4.1 +/− 0.4 days. After 5 days of storage, platelet concentrates showed a mean pH of 7.21 +/− 0.20 when prepared immediately after phlebotomy, and of 7.22 +/− 0.15 when prepared after an 8-hour delay. Mean morphology scores were 280 +/− 33 and 302 +/− 27 for platelets from units prepared immediately after phlebotomy or after a holding period of 8 hours, respectively. Platelets underwent synergistic aggregation after 5 days of storage, independent of the length of time that the units of whole blood were held prior to centrifugation. These studies indicate that platelet concentrates prepared from units of whole blood held initially for 8 hours can be stored for 5 days at 20 to 24 degrees C and survive satisfactorily in vivo and retain in vitro characteristics.     

机采血小板悬液在保存过程中凝血因子Ⅷ、Ⅸ生物活性的变化

为了研究机采血小板悬液22℃振荡保存不同时间的凝血因子Ⅷ、Ⅸ生物活性的变化,采用SYSMEXCA-1500型全自动血凝仪对用CS-3000plus血细胞分离机采集的18份血小板悬液于22℃振荡保存条件下,测定0、12、24、48、72、96、120小时7个时间段的FⅧ∶C和FⅨ∶C的活性。结果表明:机采血小板FⅧ∶C在0时活性为(100.51±44.02)%,保存12-120小时,其活性衰减了10%-40%;FⅨ∶C在0时活性为(120.93±20.50)%,保存24-120小时,其活性衰减了10%-35%。结论:机采血小板悬液于22℃振荡保存时凝血因子Ⅷ、Ⅸ仍保持有较高的生物学活性。     

In vivo survival of apheresis RBCs, frozen with 40-percent (wt/vol) glycerol, deglycerolized in the ACP 215, and stored at 4°C in AS-3 for up to 21 days

BACKGROUND: The FDA has approved the storage of frozen RBCs at -80 degrees C for 10 years and the postwash storage at 4 degrees C for no more than 24 hours. The 4 degrees C postwash storage period is limited to 24 hours, because the current deglycerolization systems are functionally open systems. STUDY DESIGN AND METHODS: Two units of RBCs were collected from each of 13 healthy male volunteers. The RBCs were collected in CP2D by the FDA-approved protocol for an automated apheresis device (MCS, LN8150, Haemonetics) and were stored at 4 degrees C in AS-3 for 6 days. Using a single disposable glycerolization set in an automated, functionally closed system (ACP 215, Haemonetics) each unit was transferred to a 1000-mL PVC plastic bag and glycerolized to a concentration of 40-percent (wt/vol) glycerol and frozen at -80 degrees C. A single disposable deglycerolization set in the ACP 215 was used to deglycerolize the 2 units from the same donor. The deglycerolized RBCs were stored at 4 degrees C in AS-3 for as long as 21 days. RESULTS: The mean +/- SD freeze-thaw-wash recovery value was 89.4 +/- 3 percent. The residual hemolysis in the RBCs stored at 4 degrees C in AS-3 for 21 days after deglycerolization was 0.9 +/- 0.2 percent, and the units were negative for both aerobic and anaerobic bacteria. The mean Nageotte WBC count was 9 x 10(6) per unit. When the deglycerolized RBCs were given as an autologous transfusion after storage at 4 degrees C in AS-3 for the 7- to 18-day period, the mean +/- SD 24-hour posttransfusion survival was 77 +/- 7 percent, and the index of therapeutic effectiveness was 69 +/- 8 percent. CONCLUSION: Two units of human RBCs collected from a single donor by apheresis in the MCS using an LN8150 set can be glycerolized sequentially with a single disposable set and deglycerolized sequentially with another single disposable set in the ACP 215. The previously frozen RBCs stored in AS-3 for 7 to 18 days at 4 degrees C had acceptable hemolysis and an acceptable mean 24-hour posttransfusion survival value and index of therapeutic effectiveness.     

Effect of rejuvenation and frozen storage on 42-day-old AS-1 RBCs

BACKGROUND: The FDA has approved a 42-day storage period for RBCs stored in ADSOL (AS-1). This study was undertaken to provide data for the FDA about the feasibility of salvaging AS-1 RBCs at the end of their storage period by rejuvenation and freezing. STUDY DESIGN AND METHOD: The investigation, consisting of a study (n = 10) and control (n = 6) arm, was carried out in two centers. In both centers, eight healthy volunteers donated a unit (450 mL) of whole blood. The RBC concentrates were stored at 4 degrees C in AS-1 for 42 days. The study units were rejuvenated, whereas the control units were not. All units were stored frozen at -80 degrees C, then deglycerolized and kept for an additional 24 hours at 4 degrees C. RESULTS: After the 42-day storage period, ATP had declined to 62 percent of the original value, 2,3 DPG was zero, and MCV was significantly larger than that of fresh RBCS: Following rejuvenation and deglycerolization, the mean ATP level was 141 percent, the mean 2,3 DPG level was 109 percent, and the MCV was normal. The freeze-thaw-wash recovery of the rejuvenated and nonrejuvenated RBCs was similar, 88.4 and 84.0 percent, respectively. There was no difference in hypoxanthine, inosine, and uric acid levels in the rejuvenated and nonrejuvenated units, which indicated that the chemicals in the rejuvenation solution and their by-products had been removed during processing. In both centers, the mean 24-hour survival of rejuvenated, deglycerolized RBCs exceeded 75 percent, whereas that of nonrejuvenated RBCs did not. The long-term survival rates of viable study and control RBCs were similar. CONCLUSION: Forty-two-day-old AS-1 RBCs that have been rejuvenated and then frozen have more than 75 percent viability and normal oxygen delivery function. Rejuvenation of RBCs does not introduce additional safety hazards to blood transfusion.     

Coagulation parameters in apheresis and leukodepleted whole-blood plasma during storage

BACKGROUND: Thawing fresh-frozen plasma (FFP) may cause delay in delivery, and one approach to circumvent this is to store plasma at +4 degrees C. Thawed plasma is commonly discarded after a few days of storage, owing to the assumption that coagulation factor activity decreases to clinically unacceptable levels. STUDY DESIGN AND METHODS: Eighteen apheresis plasma (AP) units were collected from blood donors. The collected plasma was divided into two equal parts: one part frozen at -74 degrees C as FFP and one part stored at +4 degrees C as fresh liquid plasma (FLP). Thirty-nine units of whole blood (WB) were collected from blood donors and leukodepleted by inline filtration, followed by plasma separation. Twenty plasma units were frozen at -74 degrees C as FFP and 19 plasma units were stored at +4 degrees C as FLP for 28 days. Plasma aliquots were collected before freezing and immediately after thawing FFP and before and during storage of FLP at Days 14 and 28. Factor (F)V, FVIII, D-dimers, and C1-esterase inhibitor levels were assessed. RESULTS: No significant differences in coagulation factor levels were assessed between FLP prepared from AP and FLP prepared from WB. FV and FVIII levels decreased on average 25 and 50 percent, respectively, at Day 14 of storage. C1-esterase inhibitor and D-dimers levels were not affected. CONCLUSION: Leukodepleted apheresis and WB plasma stored for 14 days retain sufficient levels of FV and FVIII activity for maintenance of normal hemostasis and could therefore be considered useful in selected clinical situations.     

Survival of Borrelia burgdorferi in blood products

The incidence of Lyme disease is rapidly increasing in the United States. To assess the potential of transmission of the disease through blood transfusion, we studied the survival of Borrelia burgdorferi in blood products under blood bank storage conditions. Two units of whole blood, separated into red cells (RBCs), fresh-frozen plasma (FFP), and platelet concentrates (PCs), were inoculated with B. burgdorferi (strain B31) in concentrations of approximately 3000 organisms per mL of RBCs and FFP and 200 organisms per mL of PCs. Products were then stored under blood banking conditions and sampled at several storage times. The viability of the spirochete in blood components was determined by darkfield microscopic examination of cultures in modified Kelly's medium. The organism was shown to survive in RBCs (4 degrees C) and FFP (below -18 degrees C) for 45 days and in PCs (20-24 degrees C) for 6 days. The results of this study do not exclude the possibility of transmission of Lyme disease through blood transfusion.     

A multicenter study of in vitro and in vivo values in human RBCs frozen with 40-percent (wt/vol) glycerol and stored after deglycerolization for 15 days at 4°C in AS-3: assessment of RBC processing in the ACP 215

BACKGROUND: The FDA has approved the storage of frozen RBCs at -80 degrees C for 10 years. After deglycerolization, the RBCs can be stored at 4 degrees C for no more than 24 hours, because open systems are currently being used. Five laboratories have been evaluating an automated, functionally closed system (ACP 215, Haemonetics) for both the glycerolization and deglycerolization processes. STUDY DESIGN AND METHODS: Studies were performed at three military sites and two civilian sites. Each site performed in vitro testing of 20 units of RBCs. In addition, one military site and two civilian sites conducted autologous transfusion studies on ten units of previously frozen, deglycerolized RBCs that had been stored at 4 degrees C in AS-3 for 15 days. At one of the civilian sites, 10 volunteers received autologous transfusions on two occasions in a randomized manner, once with previously frozen RBCs that had been stored at 4 degrees C in AS-3 for 15 days after deglycerolization and once with liquid-preserved RBCs that had been stored at 4 degrees C in AS-1 for 42 days. RESULTS: The mean +/- SD in vitro freeze-thaw-wash recovery value was 87 +/- 5 percent; the mean +/- SD supernatant osmolality on the day of deglycerolization was 297 +/- 5 mOsm per kg of H(2)O, and the mean +/- SD percentage of hemolysis after storage at 4 degrees C in AS-3 for 15 days was 0.60 +/- 0.2 percent. The paired data from the study of 10 persons at the civilian site showed a mean +/- SD 24-hour posttransfusion survival of 76 +/- 6 percent for RBCs that had been stored at 4 degrees C for 15 days after deglycerolization and 72 +/- 5 percent for RBCs stored at 4 degrees C in AS-1 for 42 days. At the three sites at which 24-hour posttransfusion survival values were measured by three double-label procedures, a mean +/- SD 24-hour posttransfusion survival of 77 +/- 9 percent was observed for 36 autologous transfusions to 12 females and 24 males of previously frozen RBCs that had been stored at 4 degrees C in AS-3 for 15 days after deglycerolization. CONCLUSION: The multicenter study showed the acceptable quality of RBCs that were glycerolized and deglycerolized in the automated ACP 215 instrument and stored in AS-3 at 4 degrees C for 15 days.     

Phosphate ion exchange resin used in the liquid preservation of baboon red cells

Baboon whole blood, collected in 14 percent citrate-phosphate-dextrose anticoagulant solution in plastic bags was stored in 100-ml aliquots at 4 degrees C for 28 days in the presence or absence of 0.75 grams of phosphate anion exchange resin. In vitro and in vivo measurements after autologous transfusions were made to determine whether the phosphate anion exchange resin had any beneficial effect on the blood during storage. The in vitro measurements of red cell 2,3-diphosphoglycerate and P50 were higher throughout the 28 days of storage at 4 degrees C in the blood stored in the phosphate anion exchange resin. After autologous transfusions in six baboons of red cells prepared from whole blood stored at 4 degrees C for 21 days, the 24-hour posttransfusion survival values were 86 +/- 6 percent (mean +/- SD) in the presence of resin and 83 +/- 6 percent in the absence of resin. In five other baboons, red cells prepared from 28-day-old blood showed a mean 24-hour posttransfusion survival of 82 +/- 4 percent in the presence of resin and 75 +/- 4 percent in the absence of resin. The addition of a phosphate anion exchange resin to the citrate-phosphate-dextrose anticoagulant provided better maintenance of red cell 2,3-diphosphoglycerate concentrations and P50 levels during storage of whole blood at 4 degrees C, and red cells prepared from whole blood stored in this solution had better oxygen transport function than red cells prepared from blood stored without resin. Red cell adenosine triphosphate concentrations and 24-hour posttransfusion survival values were similar whether or not the anticoagulant contained resin.     

Accumulation of DI-2-Ethylhexyl Phthalate (DEHP) in Whole Blood, Platelet Concentrates, and Platelet-Poor Plasma

The concentration of the plasticizer, di-2-ethylhexyl phthalate (DEHP), in the plasma was measured after storage of the whole blood in polyvinylchloride plastic bags at 4 C for up to 38 days in either ACD or CPD. The plasma from ACD-stored whole blood contained more DEHP than that from CPD-stored whole blood. The continuous-flow centrifugation washing procedures removed about 98 per cent of the DEHP from ACD whole blood stored for 33 days at 4 C.
DEHP was assayed in the platelet-rich plasma, platelet-poor plasma, supernatant of the platelet concentrate, washed platelets, and washed red blood cells prepared from CPD whole blood. Very little DEHP was found in the washed red blood cells and platelets, a small amount was found in the platelet-poor plasma, and a large amount in the supernatant of the platelet concentrate. A greater amount of DEHP accumulated in the platelet concentrates that were stored at 22 C than in those stored at 4 C. When platelet concentrates from CPD whole blood were stored at 22 C for 72 hours, the amount of DEHP was about four times that observed after 4 C storage for the same length of time.