Successful storage of RBCs for 10 weeks in a new additive solution

BACKGROUND: The effect of storing packed RBCs suspended in 300 mL of an alkaline, experimental additive solution (EAS 64) was explored. STUDY DESIGN AND METHODS: RBC units prepared from blood collected from healthy donors into CPD were WBC reduced and stored for 10 weeks under blood bank conditions after the addition of 300 mL of EAS 64 (adenine, 2 mM:; dextrose, 50 mM:; mannitol, 20 mM:; NaCl, 75 mM:; Na(2)HPO(4), 9 mM:). For comparison, non-WBC-reduced units from the same donors were stored in a different additive solution (AS-1, Baxter Healthcare) for 6 weeks. Standard methods were used for the in vitro assays. The 24-hour in vivo recoveries were measured by using (51)Cr- and (99m)Tc-labeled RBCs. RESULTS: Mean recovery in the EAS 64 units after 10 weeks was 84 +/- 8 percent, the same as in the AS-1 units stored for 6 weeks. For EAS 64 and AS-1 units, respectively, the ATP of the RBCs was 85 percent and 64 percent of the initial value, hemolysis was 0.43 percent and 0.63 percent, supernatant potassium was 24 mEq per L and 44 mEq per L, and the morphologic index was 98 and 71. CONCLUSION: RBCs suspended in 300 mL of EAS 64 can be stored satisfactorily for 10 weeks. Longer RBC storage should reduce outdating, increase availability of transfusions in remote locations, and improve the efficiency of autologous donor programs.     

RBC storage for 11 weeks

BACKGROUND: Increasing the length of RBC storage can increase both RBC availability and quality. This work addresses 11-week RBC storage in experimental ASs (EASs). STUDY DESIGN AND METHODS: Three studies were performed. In the first, 24-hour in vivo recovery of (51)Cr-labeled autologous RBCs was measured in nine volunteers after storage of their RBCs for 11 weeks in EAS 67. In the second study, 4 units of blood were divided and stored in aliquots with an EAS containing 0, 15, 30, or 45 mmol per L of mannitol; then hemolysis, RBC morphology, and microvesicle protein were measured. In the third study, 6 full units were stored for 12 weeks in the EAS containing 30 mmol per L of mannitol, with weekly sampling for morphologic and biochemical measures of RBC quality. RESULTS: RBCs stored for 11 weeks in EAS-67 had a mean 24-hour in vivo recovery of 79 +/- 5 percent, but the hemolysis was 1.35 +/- 0.68 percent. Increasing mannitol content of the EAS reduced hemolysis but increased microvesiculation. EAS-76, with 30 mmol per L of mannitol allowed 11-week storage with 0.48 +/- 0.10 percent hemolysis at 11 weeks and 0.62 +/- 0.14 percent hemolysis at 12 weeks. CONCLUSION: It is possible to store RBCs for 11 weeks in EAS with greater than 75 percent recovery and less than 1 percent hemolysis.     

Successful storage of RBCs for 9 weeks in a new additive solution

BACKGROUND: This study explored the effect of storing packed RBCs suspended in 200 mL of an alkaline, hypotonic, experimental additive solution (EAS 61). STUDY DESIGN AND METHODS: Packed RBC units prepared from RBCs collected from healthy donors in CPD were stored for 8 (n = 10) and 9 (n = 10) weeks under blood bank conditions after the addition of 200 mL of EAS 61 (adenine, 2 mM:; dextrose, 110 mM:; mannitol, 55 mM:; NaCl, 26 mM:; Na(2)HPO(4), 12 mM:). Standard methods were used for in vitro assays. The 24-hour in vivo autologous recoveries were measured with (51)Cr. RESULTS: Mean +/- SD recoveries at 8 and 9 weeks were 81 +/- 7 and 77 +/- 7 percent. After 9 weeks, the ATP of the RBCs was 81 percent of the initial value, hemolysis was 0.35 percent, supernatant potassium was 46 mEq per L, and the morphologic index was 94.1. CONCLUSION: Packed RBCs suspended in 200 mL of EAS 61 can be stored satisfactorily for 9 weeks. Longer RBC storage should reduce outdating, increase availability of transfusions in remote locations, and improve the efficiency of autologous donor programs.     

A new apheresis procedure for the preparation of high-quality red cells and plasma

BACKGROUND: Multicomponent apheresis is an alternative way of preparing blood components that avoids the delay between collection and separation seen with standard whole-blood techniques. STUDY DESIGN AND METHODS: An apheresis device has been modified to facilitate the combined collection of a unit (250 mL) of red cells (RBCs) and a high-volume unit (475 mL) of plasma. The procedure, using 8-percent ACD-A, has been tested in two European blood centers. Each center performed 20 procedures for in vitro evaluation of collected RBCs and plasma and 10 procedures for evaluation of in vivo RBC recovery. All RBCs were white cell reduced by filtration. One-half of the RBC units were stored in the additive solution Adsol and one-half in another such solution (Erythro-Sol). RESULTS: The target volumes of RBCs and plasma were obtained in 27 minutes (range, 20-44 min) by using three to six cycles in a single-needle procedure. Saline (275 mL) was used to replace fluid volume withdrawn in excess of standard whole-blood donation. No side effects occurred, with the exception of minor signs of hypocalcemia. RBC ATP was well maintained (>65% at Day 42) during storage; 2,3-DPG was less well maintained, with virtually none remaining at Day 21 in either Adsol or Erythro-Sol. The RBC in vivo recoveries, after 42 days of storage at 4+/-2 degrees C determined by the single-label method, were 86.7+/-7.2 percent (Erythro-Sol) and 84.4+/-8.1 percent (Adsol). Mean plasma factor VIII levels were >100 percent in all test groups. CONCLUSION: A novel automated technique for the simultaneous collection and preparation of RBCs and plasma has been evaluated. The apheresis procedure was acceptable and well tolerated by donors, and it resulted in high-quality blood components. Further optimization of the system should yield a practicable component suitable for routine use in blood banks.     

Evaluation of a concurrent multicomponent collection system for the collection and storage of WBC-reduced RBC apheresis concentrates

BACKGROUND: Multicomponent apheresis procedures offer the possibility of collecting blood components that are standardized, as compared to those available with whole-blood donations. A new separator program for the concurrent collection of RBCs, platelets, and plasma (Amicus, Baxter Healthcare) was evaluated. STUDY DESIGN AND METHODS: Apheresis donors (n = 47) underwent concurrent collection of RBCs, platelets, and plasma by use of the single-needle procedure of the Amicus blood cell separator. A standardized RBC volume (100% Hct) of 200 mL was targeted with either 1 or 2 platelet concentrate units, depending on the donor's predonation characteristics. After collection, the RBC component was sterilely connected to an RBC collection set (Amicus) to allow for the addition of 100 mL of saline-adenine-glucose-mannitol preservative solution and WBC reduction at either ambient temperature or 4 degrees C. The RBC units were subsequently stored at 2 to 6 degrees C for 42 days, and the following in vitro measures were evaluated over the storage period: blood cell counts including Hct and total Hb, plasma Hb, potassium, pH, ATP, and 2,3 DPG. RESULTS: Procedure time averaged 74 +/- 9 minutes, and no adverse events were reported. The absolute RBC volume collected averaged 198 +/- 11 mL with an average Hct value of 83 +/- 2 percent. After filtration, the Hb content averaged 58.2 +/- 2.4 g per unit and residual WBCs averaged 0.038 +/- 0.015 x 10(6) per unit. Day 42 results showed that all units had on average more than 70-percent ATP maintenance, and all of the units had less than 0.8 percent he-molysis. All units had pH values higher than 6.5 on Day 42. CONCLUSION: The concurrent multicomponent collection system (Amicus) can reliably collect a standardized RBC unit of good quality. In vitro testing of the RBCs collected and stored for 42 days met the Council of Europe criteria for transfusion.     

Evaluation of an automated system for the collection of packed RBCs, platelets, and plasma

BACKGROUND: This study evaluated the quality of WBC-reduced platelets, RBCs, and plasma collected on a new system (Trima, Gambro BCT) designed to automate the collection of all blood components. The study also evaluated donor safety and suitability of these components for transfusion. STUDY DESIGN AND METHODS: In Phase I, the quality of the components collected on the new system was evaluated by standard in vitro and in vivo testing methods. Results were compared to those from control components collected by currently approved standard methods. In Phase II, additional collections were performed to evaluate the acceptability of the new system and the safety of platelets collected. RESULTS: In vivo 24-hour RBC recovery was 76.8 +/- 3.1 percent for the test RBC units and 77.1 +/- 4.4 percent recovery for whole-blood (control) RBCs. The differences between test and control platelet results in the in vivo and in vitro assays were not clinically significant. Plasma clotting factors and fibrinogen levels met international standards. The system was well accepted by donors, and no major adverse donor reactions were reported for the 68 procedures performed. No problems were reported with transfusing the blood components collected. CONCLUSION: Blood components collected with the Trima are equivalent to currently available components, and they meet the applicable regulatory standards. This system provides consistent, standardized components with predictable yields. It provides the option of fully automating the collection of all blood components.     

Processing of major ABO-incompatible bone marrow for transplantation by using dextran sedimentation

BACKGROUND: Various open and semi-closed methods are used for red cell (RBC) depletion and hematopoietic progenitor cell (HPC) enrichment of bone marrow (BM) in vitro, but with variable efficacy. A simple, efficient, and safe method using dextran 110k was developed. STUDY DESIGN AND METHODS: An equal volume of 4.5-percent dextran was applied to major ABO-incompatible BM in transfer bags and sedimentation was allowed for 30 minutes. RBCs, nucleated cells (NCs), and mononuclear cells (MNCs) from BM allografts before and after dextran sedimentation (DS) were counted. Flow cytometry, short-term cultures, and long-term cultures were performed to assay the respective recovery of CD34+ cells, colony-forming units (CFUs), and long-term culture-initiating cells (LTC-ICs). RESULTS: Sixteen BM collections were processed.The mean volume was 666 mL (range, 189-1355 mL).The mean +/-1 SD post-DS NC, MNC, CD34+ cell, and CFU counts per kg of the recipient's body weight were 4.11 +/-1.74 x 10(8), 8.98 +/- 3.68 x 10(7), 2.90 +/- 1.95 x 10(6), and 2.03 +/- 2.01 x 10(5), respectively, with the corresponding post-DS recovery being 90.6 percent, 90 percent, 92.4 percent, and 100.8 percent. The numbers of LTC-ICs in cultures (up to 12 weeks) of pre-DS and post-DS samples of five BM allografts were comparable (p = 0.91). Residual RBCs were 5.1 +/- 4.6 (0.1-14) mL with depletion of 96.5 +/- 3.2 percent. There was no significant difference in the mean absolute RBC count in post-DS BM allografts and in four ficoll-treated BM allografts (8.09 x 10(10) vs. 4.9 x 10(9); p = 0.206) and in eight major ABO-incompatible peripheral blood HPC collections (8.09 x 10(10) vs. 9.81 x 10(10); p = 0.87). No posttransplant hemolysis was encountered. Engraftment occurred at 22 +/- 7 days, which is similar to that of four transplants with ficoll-treated BM allografts (22 +/- 9; p = 0.611) and 54 unprocessed BM allografts (19 +/- 6; p = 0.129). CONCLUSION: DS is an efficient method of depleting RBCs in major ABO-incompatible BM allografts without significant loss of HPCs.     

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.     

Quality and clinical response to transfusion of prestorage white cell-reduced apheresis platelets prepared by use of an in-line white cell-reduction system

BACKGROUND: This study evaluated the quality and clinical effectiveness of white cell (WBC)-reduced apheresis platelets collected by the use of a new technology, fluidized particle-bed separation. STUDY DESIGN AND METHODS: In phase 1, six suitable donors underwent two separate plateletpheresis procedures on one occasion, each separated by less than 10 minutes. In random order, a control unit was collected with the COBE Spectra and a test unit with the Spectra Leukocyte-Reduction System (LRS). The quality of apheresis platelet components was assessed by an in vitro test panel, and residual WBCs were counted by Nageotte chamber and flow cytometric methods. For the in vivo studies, the test and control units were randomly labeled with either 51Cr or 111In at the end of storage and transfused simultaneously to the donor. Samples were taken for calculation of platelet survival and recovery. In phase II, 109 thrombocytopenic patients were given platelets collected by use of the Spectra LRS. RESULTS: Test platelets had significantly fewer residual WBCs (median 7.6 x 10(4)) than control platelets (median 3.9 x 10(5)), with equivalent in vitro function values. Test and control platelets had similar recovery and survival. Transfused platelets collected by use of the LRS achieved a mean 1-hour corrected-count increment of 19.3. CONCLUSION: The LRS collects platelet components with significantly lower WBC contamination without adverse effects on the function or in vivo survival of the platelets.     

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.     

WBC-reduced platelet concentrates from pooled buffy coats in additive solution: an evaluation of in vitro and in vivo measures

BACKGROUND: The use of a platelet additive solution (PAS-II, Baxter) may have benefits over plasma for storage of platelets. It was the aim of this study to develop a method to produce WBC-reduced platelet concentrates (PCs) in PAS-II with >240 x 10(9) platelets and <1 x 10(6) WBCs per unit, which can be stored for 5 days at pH >6.8 and that will give sufficient platelet increments after transfusion: a 1-hour CCI of >7.5 and a 20-hour CCI of >2.5. STUDY DESIGN AND METHODS: PCs were made from five pooled buffy coats and 250 g of PAS-II. After centrifugation the PCs were WBC-reduced with a filter (Autostop BC, Pall Biomedical) and stored in a 1000-mL polyolefin container. CCIs were assessed in stable hemato-oncologic patients after 5-day old PCs were transfused. RESULTS: Routinely produced PCs contained a median of 310 x 10(9) platelets (n = 5,363) with 3.5 percent containing <240 x 10(9) platelets, in a median volume of 320 mL (n = 11,834). The median number of WBCs was <0.03 x 10(6) (n = 694). The WBC count exceeded 1 x 10(6) in three PCs, but it was always <5 x 10(6), giving 99-percent confidence that more than 99.5 percent of the units will contain <1 x 10(6) WBCs. The pH remained >6.8 on Day 8, provided the concentration was below 1.1 x 10(9) platelets per mL (n = 32). After 28 transfusions in 28 patients, the 1-hour CCI was 12.6 +/- 4.3 (mean +/- SD, with 2/28 CCIs <7.5) and the 20-hour CCI was 8.9 +/- 5.6 (with 4/28 CCIs <2.5). Limitations of this study include the absence of a control group of patients receiving platelets stored in plasma and of in vivo radiolabeled survival studies, but a comparison of these data with previously published data suggested that the in vivo survival of platelets stored in PAS-II is less than that of platelets stored in plasma. CONCLUSION: The WBC-reduced PCs conformed to specifications. These WBC-reduced PCs could be stored at least 5 days with maintenance of pH, and they gave sufficient increments after transfusion to patients.     

Evaluation of a whole-blood WBC-reduction filter that saves platelets: in vitro studies

BACKGROUND: In this study, a new WBC-reduction in-line filter that removes WBCs but not platelets was evaluated. Three WBC-reduced blood components were prepared: RBCs, plasma, and platelet concentrates (PCs). STUDY DESIGN AND METHODS: Whole-blood components (n = 30) were filtered within 2 to 4 hours after collection and then were centrifuged and separated into RBCs, plasma, and WBC-reduced buffy coat. Saline-adenine-glucose-mannitol solution was added to the RBCS: The WBC-reduced buffy coats were stored overnight; on the following day, PCs were prepared from pooled WBC-reduced buffy coats and stored in a medium composed of approximately 35 percent CPD plasma and 65 percent platelet additive solution (T-Sol, Baxter). The WBC-reduction capacity of the filter, the recovery of cells after filtration, and the in vitro storage of RBCs (n = 10) and platelets (n = 6) were evaluated. RESULTS: Mean and maximum WBC counts after filtration were 0.08 x 10(6) and 0.3 x 10(6), respectively, per filtered whole-blood unit. Recovery of RBCs (mean values) after filtration was 90 percent in whole-blood components and 73 percent in RBCS: Recovery of platelets (mean values) was 81 percent after filtration and 66 percent in PCS: The in vitro storage study of RBCs showed results comparable with previously published data, except for a lower degree of hemolysis. In the in vitro platelet storage study, results were compared with those of standard preparations. In all essentials, similar results were found. CONCLUSION: The results of the present study suggest that effective WBC reduction meets current standards and satisfactory recovery after filtration. The storage characteristics for RBCs and PCs are similar to those of standard preparations. Use of a whole-blood in-line filter to save platelets is a new option for whole-blood processing, which may simplify WBC reduction and blood component preparation, as well as reduce costs in the future.     

The viability of autologous human red cells stored in additive solution 5 and exposed to 25°C for 24 hours

BACKGROUND: No data exist on the viability of red cells (RBCs) stored in modern additive solution systems and allowed to warm above 10 degrees C. STUDY DESIGN AND METHODS: In a randomized crossover study, 3 units of blood were collected at least 8 weeks apart from 11 volunteer donors and stored in additive solution 5 (AS-5). Of 3 units from each volunteer, 1 was stored for 6 weeks at 4 degrees C, 1 for 5 weeks at 4 degrees C except for 24 hours at 25 degrees C on Day 14, and 1 for 5 weeks at 4 degrees C except for 24 hours at 25 degrees C on Day 28. Units were sampled periodically during storage; at the end of storage, viability was measured by the 99mTc/51 Cr double-label method. RESULTS: RBC viability was not significantly different in the storage protocols. Less than 1 percent of stored cells hemolyzed. RBC ATP concentrations at the end of storage correlated with viability and were approximately equal in the warmed units after 30 days' storage and the conventionally stored units after 42 days. CONCLUSIONS: The data suggest that RBCs stored in AS-5 and allowed to warm to 25 degrees C for 24 hours lose about 12 days of their shelf life.     

In vitro collection and posttransfusion engraftment characteristics of MNCs obtained by using a new separator for autologous PBPC transplantation

BACKGROUND: A clinical study was performed to evaluate the peripheral blood progenitor cell (PBPC) collection, transfusion, and engraftment characteristics associated with use of a blood cell separator (Amicus, Baxter Healthcare). STUDY DESIGN AND METHODS: Oncology patients (n = 31) scheduled for an autologous PBPC transplant following myeloablative therapy were studied. PBPCs were mobilized by a variety of chemotherapeutic regimens and the use of G-CSF. As no prior studies evaluated whether PBPCs collected on the Amicus separator would be viable after transfusion, to ensure patient safety, PBPCs were first collected on another cell separator (CS-3000 Plus, Baxter) and stored as backup. The day after the CS-3000 Plus collections were completed, PBPC collections intended for transfusion were performed using the Amicus instrument. For each transplant, >2.5 x 10(6) CD34+ PBPCs per kg of body weight were transfused. RESULTS: Clinical data collected on the donors immediately before and after PBPC collection with the Amicus device were comparable to donor data similarly obtained for the CS-3000 Plus collections. While the number of CD34+ cells and the RBC volume in the collected products were equivalent for the two devices, the platelet content of the Amicus collections was significantly lower than that of the CS-3000 Plus collections (4.35 x 10(10) platelets/bag vs. 6.61 x 10(10) platelets/bag, p<0.05). Collection efficiencies for CD34+ cells were 64 +/- 23 percent for the Amicus device and 43 +/- 14 percent for the CS-3000 Plus device (p<0.05). The mean time to engraftment for cells collected via the Amicus device was 8.7 +/- 0.7 days for >500 PMNs per microL and 9.7 +/- 1.5 days to attain a platelet count of >20,000 per microL-equivalent to data in the literature. No CS-3000 Plus backup cells were transfused and no serious adverse events attributable to the Amicus device were encountered. CONCLUSIONS: The mean Amicus CD34+ cell collection efficiency was better (p<0.05) than that of the CS-3000 Plus collection. Short-term engraftment was durable. The PBPCs collected with the Amicus separator are safe and effective for use for autologous transplant patients requiring PBPC rescue from high-dose myeloablative chemotherapy.     

In vivo and in vitro characteristics of double units of RBCs collected by apheresis with a single in-line WBC-reduction filter

BACKGROUND: A novel apheresis procedure for a blood separator (MCS+, Haemonetics) enables the collection of 2 WBC-reduced RBC units in a single donation by using one disposable set with one in-line WBC-reduction filter (RC2H, Pall Corp.). The objective of this study was to evaluate the filtration performance in connection with different prefiltration RBC storage conditions and with the in vitro and in vivo storage quality of the filtered units. STUDY DESIGN AND METHODS: Sixty-six 2-unit RBC collection and gravity-filtration procedures were completed at three sites, resulting in 132 RBC units. Filtration of the double RBC units was performed at room temperature (RT) within 8 hours of collection (n = 36) and under refrigeration (1-6 degrees C) for up to 24 hours (n = 10) and 72 hours (n = 20) before filtration. RBC quality was compared to that of nonfiltered apheresis RBC units (n = 10). RESULTS: Median filtration time was 6.5 and 14 minutes for units stored at RT and under refrigeration, respectively. All 132 RBC units had residual WBC counts <0.4 x 10(6). The refrigerated units showed a greater mean log reduction in WBCs: 5.06 +/- 0.16 (24 hour) and 4.74 +/- 0.48 (72 hour), respectively, than did RT units: 4.47 +/- 0.28 (p<0.05). RBC loss was less than 12 percent in all cases (mean, 7.8 +/- 1.8%). Minimal differences in volume were observed between the paired RBC units. In vitro RBC storage characteristics of the filtered units were as expected and similar to those of the nonfiltered units. For RBC units held at RT (n = 24), the mean in vivo 24-hour recovery was 81.8 +/- 8.4 percent (double-label). CONCLUSION: Satisfactory filter performance in terms of WBC removal and RBC loss was observed with all 66 procedures, irrespective of storage conditions before filtration.     

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.     

Comparison of two sterile connection devices and the effect of sterile connections on blood component quality

BACKGROUND: Sterile connection devices (SCDs) are used to connect pieces of polyvinylchloride tubing between blood bag systems. After observing a slight decrease in inner diameter of tubing welded with the CompoDock S2 SCD, the effect of welded tubing on storage characteristics of white blood cell (WBC)-reduced red blood cells (RBCs) and platelet (PLT) concentrates was studied. Welds were made with Terumo SCD (T-SCD) or CompoDock S2, and unwelded tubing served as reference. STUDY DESIGN AND METHODS: Three WBC-reduced RBC units or 3 PLT concentrates were pooled and divided to prevent donor-dependent differences. The units were transferred 10 times over (1) tubing with a T-SCD weld, (2) a CompoDock S2 weld, or (3) unwelded tubing. RBCs were stored for 42 days and free hemoglobin (Hb) was measured; PLT concentrates were stored for 8 days and CD62P expression was measured, as markers for blood component quality (n = 10 paired experiments). RESULTS: WBC-reduced RBC units had similar hemolysis at the end of storage: 0.47 +/- 0.28, 0.47 +/- 0.35, and 0.49 +/- 0.38 percent of total Hb, for tubing with a T-SCD weld, a CompoDock S2 weld, or no weld, respectively (not significant). CD62P expression of stored WBC-reduced PLT concentrates was not significantly different between the groups: 20.3 +/- 5.1, 19.8 +/- 5.1, and 22.3 +/- 9.8 percent for tubing with a T-SCD weld, a CompoDock S2 weld, or no weld, respectively. CONCLUSION: The quality of blood components, measured as RBC hemolysis and platelet CD62P expression, is not adversely affected by the presence of a sterile connection in the tubing, made by either the CompoDock S2 or the T-SCD.     

The role of electrolytes and pH in RBC ASs

BACKGROUND: Experimental additive solutions (EASs) containing saline, adenine, glucose, mannitol and disodium phosphate can support RBCs for 9 or 10 weeks if used in 200- or 300-mL volumes. The effects of variations in the electrolyte composition and volume of EASs were explored. STUDY DESIGN AND METHODS: In three four-arm studies, 24 RBC units were pooled in groups of 4 and realiquoted as test units to ensure that all donors were equally represented in each study arm. In Study 1, units were stored for 11 weeks in EAS containing 0, 10, 20, or 30 mmol per L of sodium bicarbonate. In Study 2, units were stored for 9 weeks in EAS containing 26, 50, 100, or 150 mmol per L of sodium chloride. In Study 3, units were stored in 100 or 200 mL of AS-3 or EAS-61. RBC ATP concentrations and hemolysis were measured weekly. RESULTS: Increasing the sodium bicarbonate content of EASs increased the pH throughout storage and increased RBC ATP concentrations in the later phases of storage, but it had no effect on hemolysis. Increased sodium chloride content of EASs led to lower RBC ATP concentrations and increased hemolysis. In EAS-61, RBC ATP concentrations were increased throughout storage, and hemolysis was lower than that of RBCs stored in AS-3. CONCLUSION: RBC ATP synthesis is highly dependent on the pH of the AS. Hemolysis is affected by the salt content and volume of the AS.     

Automated collection of double red blood cell units with a variable-volume separation chamber

BACKGROUND: Automated collection of blood components offers multiple advantages and has prompted development of portable devices. This study sought to document the biochemical and hematologic properties and in vivo recovery of red cells (RBCs) collected via a new device that employed a variable-volume centrifugal separation chamber. STUDY DESIGN AND METHODS: Normal subjects (n = 153) donated 2 units of RBCs via an automated blood collection system (Cymbal, Haemonetics). Procedures were conducted with wall outlet power (n = 49) or the device's battery source (n = 104). Units were collected with or without leukoreduction filtration and were stored in AS-3 for 42 days. The units were assessed via standard biochemical and hematologic tests before and after storage, and 24 leukoreduced (LR) and 24 non-LR RBCs were radiolabeled on Day 42 with Na(2)(51)CrO(4) for autologous return to determine recovery at 24 hours with concomitant determination of RBC volume via infusion of (99m)Tc-labeled fresh RBCs. RESULTS: Two standard RBC units (targeted to contain 180 mL of RBCs plus 100 mL of AS-3) could be collected in 35.7 +/- 2.0 minutes (n = 30) or 40.3 +/- 2.7 minutes for LR RBCs (n = 92). An additional 31 collections were conducted successfully with intentional filter bypassing. RBC units contained 104 +/- 4.1 percent of their targeted volumes (170-204 mL of RBCs), and LR RBCs contained 92 percent of non-LR RBCs' hemoglobin. All LR RBCs contained less than 1 x 10(6) white blood cells. Mean hemolysis was below 0.8 percent (Day 42) for all configurations. Adenosine triphosphate was well preserved. Mean recovery was 82 +/- 4.9 percent for RBCs and 84 +/- 7.0 percent for LR RBCs. CONCLUSIONS: The Cymbal device provided quick and efficient collection of 2 RBC units with properties meeting regulatory requirements and consistent with good clinical utility.     

The effects of phosphate, pH, and AS volume on RBCs stored in saline-adenine-glucose-mannitol solutions

BACKGROUND: RBC ATP concentrations are the most important correlate of RBC viability. Tests were performed to determine whether increased AS volume, pH, and phosphate content increased stored RBC ATP concentrations. STUDY DESIGN AND METHODS: In three studies, packed RBCs were pooled in groups of 3 or 4 units and realiquoted as combined units to reduce intradonor differences. Pooled units were stored in the licensed ASs, AS-1 or AS-5, which contain saline, adenine, glucose, and mannitol (SAGM), or in experimental ASs (EASs) containing SAGM and disodium phosphate. Ten pools were stored in AS-1 at RBC concentrations equivalent to 100, 200, or 300 mL of AS. Six pools were stored in 100, 200, 300, or 400 mL volumes of EAS-61. Ten pools were stored in 100 mL of AS-5, 200 mL of EAS-61, or 300 mL of EAS-64. RBC ATP concentration and other measures of RBC metabolism and function were measured weekly. RESULTS: RBC ATP concentrations decreased sooner with storage in increasing volumes of AS-1. In EAS-61 and EAS-64, RBC ATP concentrations initially increased and stayed elevated longer with increasing AS volume. CONCLUSIONS: The addition of disodium phosphate to SAGM AS increases the RBC ATP concentrations. Reducing storage Hct appears to have a separate beneficial effect in reducing hemolysis.