A new polyvinylchloride blood bag plasticized with less-leachable phthalate ester analogue, di-n-decyl phthalate, for storage of platelets

To compare changes in platelets stored in the new di-n-decyl phthalate (DnDP)-plasticized polyvinyl chloride (PVC) bag with those in a di-(2-ethylhexyl) phthalate (DEHP)-plasticized PVC bag, single-donor apheresis platelet concentrates (PCs), 133 +/- 11 x 10(7) platelets per ml (n = 7), were stored with 94 +/- 3 ml of plasma in a new 1-liter bag with a surface area of 44 +/- 7.1 cm2 per 10(10) platelets. Oxygen and carbon dioxide gas diffusion properties of PVC-DnDP films were respectively, 1.6 and 2 times those of standard PVC-DEHP films. The amounts of DnDP leaked into the plasma of PCs were low at 0.58 +/- 0.06 mg per bag after 5-day storage, which is about one-eightieth the amount of DEHP leaked. The pH of PCs in PVC-DnDP bags amounted to 6.99 +/- 0.03 after 5-day storage, with glycolysis accelerated somewhat in the new bags. However, the platelet oxygen consumption was no different from that in the PVC-DEHP bags. Platelet aggregation and responses to hypotonic shock were significantly better in the new bags at the end of storage. Shape changes of platelets into spherical forms with dendrites were more frequently observed in PVC-DnDP bags than in PVC-DEHP bags. The study indicated that platelets stored in the new DnDP-plasticized PVC bags have retained aggregation and responses to hypotonic shock more than platelets in the PVC-DEHP bags, but spherical forms and anaerobic metabolism increased in the new bags.     

Leukocyte-poor red blood cells prepared by the addition and removal of glycerol from red blood cell concentrates stores at 4 C

Glycerol was added to and removed from red blood cells to prepare red blood cells free of white blood cells, platelets, and plasma protein. The red blood cells were stored in the primary polyvinylchloride (PVC) plastic collection bag for up to eight days. Red blood cell concentrates not treated with glycerol were washed either within four to six hours of collection or after seven days of storage at 4 C. Red blood cells mixed with glycerol were either washed immediately after addition or were equilibrated for 15 minutes at room temperature before washing. Leukocyte removal was influenced by the length of storage of the red blood cell concentrate at 4 C after collection and by the equilibration of the red blood cell-glycerol mixture prior to washing. The greatest number of leukocyte was removed when red blood cell concentrates were stored at 4 C for at least five days, mixed with glycerol solution, and the red blood cell-glycerol mixture equilibrated for 15 minutes before washing the Haemonetics Blood Processor 115. Leukocyte-poor red blood cells prepared by this procedure have been shown to be safe and effective in eliminating febrile transfusion reactions.     

Current status of additive solutions for platelets

The storage of platelets in additive solution (PAS) had lagged behind red cell concentrates, especially in North America. The partial or complete removal of anticoagulated plasma and storage of platelet concentrates in AS presents many advantages. The PAS can be formulated to optimize aerobic metabolism or decrease platelet activation, thus abrogating the platelet storage lesion and potentially improving in vivo viability. Plasma removal has been shown to reduce allergic reactions and the plasma harvested could contribute to the available plasma pool for transfusion or fractionation. PAS coupled to pathogen reduction technology results in a platelet product of equivalent hemostatic efficacy to conventionally stored platelets. Given the above, the likely future direction of platelet storage will be in new generation designer PAS with an extended shelf life and a superior safety profile to plasma stored platelets. J. Clin. Apheresis, 2012. © 2012 Wiley Periodicals, Inc.     

Cryopreservation of single-donor platelets with a reduced dimethyl sulfoxide concentration by the addition of second-messenger effectors: enhanced retention of in vitro functional activity

BACKGROUND: The potential for bacterial contamination limits the storage of platelets at 22 degrees C to 5 days. This creates an inventory problem, which could be overcome by the use of cryopreservation to allow long-term storage of platelets. It has been demonstrated that the addition to platelets of a mixture of second- messenger effectors (platelet storage solution), allows these cells to retain significant in vitro functional activity following cold storage. Analysis is needed of the ability of this second messenger effector mixture both to protect platelets during cryopreservation and to reduce the need for a cryoprotectant. STUDY DESIGN AND METHODS: Fresh single- donor platelet units (n = 8) were divided into three samples and treated with 6-percent dimethyl sulfoxide (DMSO), 2-percent DMSO or the platelet storage solution and 2-percent DMSO. The samples were placed directly into a -80 degrees C freezer and stored for 1 week, after which they were thawed and analyzed for in vitro functional activity. RESULTS: Platelets cryopreserved with the platelet storage solution and 2-percent DMSO displayed statistically higher retention of functional activity and viability–including cell number, percent of discoid cells, extent of shape change, and hypotonic shock response–than did platelets stored by the method using 6-percent DMSO. In addition, the treated platelets displayed statistically lower expression of p- selectin. The treated platelets showed no loss of cell number, > 88- percent retention of discoid morphology, and > 75-percent retention of ristocetin-induced aggregation as compared to values for these measures in fresh platelets. CONCLUSION: The use of this platelet storage solution in the cryopreservation of platelets yields a significant improvement in their postthaw in vitro recovery and allows for a reduction of the DMSO concentration from 6 to 2 percent, with superior maintenance of in vitro viability and function.     

In vivo viability of red blood cells stored in CPDA-2

The marginal viability of erythrocytes stored for 35 days as red blood cell concentrates in citrate-phosphate-dextrose-adenine-one (CPDA-1) was attributed to inadequate nutrient support with adenine and glucose. In an effort to improve the viability of red blood cells following extended storage, a new CPD-adenine and 1.4 times more glucose than CPDA-1. The efficacy of CPDA-2 was evaluated in vivo by measurement of 24-hour postinfusion recovery of 51Cr-labeled erythrocytes which had been stored as whole blood or red blood cell concentrates for 5 to 8 weeks. All red blood cell concentrates, and the whole blood units stored for 35 and 42 days, were held at room temperature for 8 hours prior to processing and/or refrigeration. CPDA-2 yielded significantly higher 51Cr survivals than CPDA-1 and exceeded the accepted criterion for anticoagulant preservative efficacy of 70 percent postinfusion survival of red blood cells after storage for a period of 42 days. Preliminary data supports possible usage to 49 days. Plasma glucose and red blood cell ATP concentration were maintained better in CPDA-2 than in CPDA-1. When compared to historical controls for CPD and CPDA-1 the data suggest that red blood cells stored in CPDA-02 will have superior viability throughout the entire storage period. CPDA-2 is a candidate to replace CPDA-1 as the anticoagulant-preservative solution of choice for red blood cell concentrates.     

Biochemical and membrane functional alterations in red cells during preparation and storage of leukocyte- and platelet-poor red cell suspensions prepared by warm-centrifuge method

A simple technique for effective removal of leukocytes and platelets from red blood cells (RBC) was reported. In that technique whole blood was incubated at 37 degrees C. In the modified procedure phosphate buffer was added to the starting RBC. Changes in red cells during preparation and storage at 4 degrees C of RBC suspensions were evaluated. Reconstituted RBC were incubated at 37 degrees C for 1 hr. After centrifugation, the lower compartment was separated with a special apparatus and diluted with 0.9% saline, which contained 78% red cells and 3% leukocytes and platelets of the starting RBC. In the incubation and subsequent preparation steps, 2,3-diphosphoglycerate levels were decreased, but not adenosine triphosphate. One-day storage caused no serious metabolic changes in red cells. Red cell osmotic fragility was not changed throughout. Stored RBC maintained sterility. The in vitro data indicate that red cells in leukocyte- and platelet-poor RBC suspensions processed with the warm-centrifuge method retained high biochemical and membrane functional abilities for 1 day.     

Platelet collection and transfusion using the fenwal CS-3000 cell separator

A prototype model of the Fenwal CS-3000 Blood Cell Separator (Deerfield, IL) was studied for plateletpheresis in 63 donors and 5 transfusions in patients. Donor effects were consistent with platelet removal and mild hemodilution. The incidence of reactions (9 of 63) was low and all were mild "citrate" type. A two-hour collection yielded 4.0 ± 0.72 × 10(11) platelets at an efficiency of 45 ± 6.9 per cent. The product had little contamination with leukocytes (0.26 ± 1.2 × 10(9) and red blood cells (hematocrit less than 1%). Morphology and pH were well preserved during 24 hours of storage. Four patients with uncomplicated aregenerative thrombocytopenia were transfused on five occasions, with a mean of 4.5 ± 0.87 × 10(11) platelets resulting in a mean platelet count increment of 55,000/microliter and dramatic reduction in template bleeding times.     

Preparation of blood components with saline-adenine-glucose-phosphate- maltose quadruple-pack system

A saline-adenine-glucose-phosphate-maltose (SAGP-maltose) quadruple-pack system was devised to improve the conventional method of preparing blood components. One of the features of this method is that high-speed centrifugation was used as the initial spin in the two-step centrifugation of whole blood. This feature permitted the removal of the buffy coat from red cells at the same time the platelets were isolated. Since the platelets were concentrated without forming a button, the damage to the platelets due to mechanical stress in preparation was minimized. Another major feature was the use of the SAGP-maltose solution. Maltose effectively prevented hemolysis during storage in a protein-poor medium for preserving red cells. Judging from the red cell adenosine triphosphate level and morphology score, it should be possible to store red cells for 35 to 42 days in SAGP-maltose preservative. In addition, approximately 25 percent more plasma was collected in the present system than in conventional multiple-pack systems.     

The effect of delayed refrigeration on red blood cells, platelet concentrates and cryoprecipitable AHF

The effect of delaying blood processing for six hours while maintaining it at ambient temperature was investigated. Blood drawn from volunteers on two occasions was processed immediately (I) or after a six-hour delay (D). The effects of the delay on the efficacy and safety of red blood cells, platelet concentrates and cryoprecipitable AHF were studied. There was a more rapid decrease in 2,3-DPG in the delayed group during 21 days of refrigerated storage. ATP levels declined at similar rates. 24-hour survival of 51CR-labeled autologous cells was slightly better (p = .05) in the (I) group but excellent for both. Total platelets, per cent recovery and pH at 72 hours were identical for both groups. All cultures were sterile. There was no difference in total AHF recovered or per cent recovery between the two groups. To increase the availability of blood components, a six-hour processing delay seems warranted.     

Current transfusion practice and need for new blood products to ensure blood supply for patients with major hemorrhage in Europe

Background

New blood products are considered for treatment of patients with major hemorrhage. The aim of this report is to describe the current transfusion practices in Europe for patients with major hemorrhage and explore the need for new or modified blood products to ensure prehospital and in-hospital blood supply.

Study Design and Method

The European Blood Alliance (EBA) Working Group on Innovation and New Blood Products' subgroup on major hemorrhage performed a survey among the EBA member states.

Results

The response rate was 58% (17 responses from 15 of the 26 EBA member states). Of these, sixteen (94%) provide massive transfusion packages (MTPs) with balanced ratio of red blood cells and plasma. Seven of the respondents included platelets from the start of treatment. Eleven (65%) provide prehospital blood products, mainly red cell concentrates or dried and/or thawed plasma with 5 days of extended storage. Two countries provide prehospital whole blood. Twelve respondents (71%) saw a need for implementation of new or modified blood components in their institution. The top three priorities were whole blood (12 of 12, 100%), dried plasma (8 of 12, 67%), and cold-stored platelets (7 of 12, 58%).

Discussion

Current national guidelines for use of blood products in patients with major hemorrhage in Europe agree on the use of balanced transfusion, however the timing and source of platelets differ. Blood products for prehospital transfusion are available in several European countries. An interest in new or modified blood products for patients with major hemorrhage was observed, especially for whole blood.     

可溶性HLA-I检测技术和贮存血中可溶性HLA-I的浓度变化观察

本研究建立可溶性人类白细胞抗原 I类 (sHLA I)检测方法 ,并探讨贮存血中sHLA I浓度变化的意义。采用ELISA双抗夹心法定量检测 6 0例正常广东人血清中sHLA I水平和 2 0例献血员成分血中sHLA I含量。结果表明 :采用本技术时 ,可溶性HLA I最低检测限为 2 .84ng ml,批内变异系数为 5 .80 % ,批间变异系数为 9.0 0 % ,回收率≥ 98 5 7% ,广东人sHLA I平均值为 (6 99.5 4± 36 0 .10 )ng ml。贮存 2 8天的RBC和随机供者血小板的sHLA I的浓度明显高于其它成分血 ,并且与成分血中残存的白细胞数和贮存时间有关。结论 :用ELISA法检测可溶性HLA I灵敏、特异、稳定 ,在临床输血过程中可考虑选择性输注含有不同浓度可溶性HLA I的成分血。     

The effect of platelets and red cells on granulocytes stored at 22 degrees C

Granulocyte concentrates contain varying numbers of platelets and red cells depending upon the method of collection. Either platelet or red cell concentrations may be as high as 2.0 × 10(12) per I. Studies were done on unwashed and washed granulocyte concentrates and on pure granulocyte suspensions with known numbers of platelets or red cells added. These suspensions or concentrates were stored for 72 hours at 22 degrees C. In both experiments, the following were measured: leukocyte and absolute granulocyte counts, dye exclusion, chemotaxis, plasma glucose, plasma pH, and osmolality. Red cell contamination did not adversely effect granulocyte storage. Platelets, however, did contribute to the functional deterioration of stored granulocytes. In the presence of high concentrations of platelets, both granulocyte dye exclusion and chemotaxis were adversely affected at 48 hours of storage. In another experiment, fresh autologous granulocytes incubated for 18 hours in hydroxyethyl starch-citrated-plasma obtained from stored granulocyte concentrates showed a progressive decrease in chemotaxis related to the age of the stored plasma. Glucose supplementation of the spent plasma maintained chemotactic activity. Contamination of granulocyte concentrates with other cells and the availability of glucose to granulocytes are variables affecting the short-term liquid storage of granulocytes at 22 degrees C.     

The Purification of Red Cells for Transfusion by Freeze Preservation and Washing

In the study of 125 units of washed, previously frozen red blood tells approximately 96.6 per cent of the leukocytes and 98.9 per cent of the platelets were removed. Less than 0.5 mg of plasma protein remained per unit. The length of storage as whole blood at 4 C prior to glycerolization of the red blood cells influenced the number of leukocytes in the washed, previously frozen red blood cells. Red blood cells frozen on the day of collection had 7.2 per cent residual white cells, whereas those stored at 4 C for 10 days or longer prior to glycerolization, freezing, and washing had 1.1 per cent residual leukocytes. When red blood cells were stored at 4 C for longer than one week, glycerolized, and washed without freezing, they had about 2 per cent residual leukocytes and platelets. The washing procedure removed at least 95 per cent of the platelets from nonfrozen and previously frozen red blood cells. Because of the removal of large numbers of leukocytes and platelets during the freeze-preserva-tion procedure and the excellent recovery of the red blood cells, washed, previously frozen glycerolized red cells are recommended for patients requiring red blood cells depleted of leukocytes, platelets, and plasma protein.     

Detection of cytomegalovirus antibody in stored blood products using latex agglutination

The detection of antibody to cytomegalovirus (CMV) in donor sera is one effective method for the prevention of posttransfusion CMV infection in seronegative recipients. A passive latex agglutination test (CMV Scan, Hynson, Wescott & Dunning, Baltimore, MD) has been shown to be an acceptable method of screening sera from the blood donor population. The procedure for this test, however, does not permit the testing of stored donor blood products. To evaluate the feasibility of testing blood components at various times during storage, the authors examined 25 CMV-positive and 25-CMV negative samples of CPDA-1 plasma from 50 units each of platelets and red cells. Plasma samples from platelets stored at 22 degrees C were tested on each of the 5 days of storage. Samples from red cell units stored at 2 to 6 degrees C were tested on storage Days 1, 2, 4, 6, 8, 10, 12, and 14. Of 250 tests done on plasma from platelet units, there were 123 true-positive and 123 true-negative results (sensitivity and specificity, 98.4%). Of 400 tests on plasma from red cell units, there were 200 true-positive and 198 true-negative results (sensitivity, 100%; specificity, 99.5%). These data show that the CMV Scan test can be used reliably to test segments of CPDA-1 plasma from platelets stored for up to 5 days and from red cells stored for up to 14 days.     

Cold storage of whole blood in an additive solution containing apoptotic and necrotic inhibitors

Background

Whole blood (WB) reigns superior to component therapy for the treatment of hemorrhagic shock on the battlefield. Though cold storage of WB offers a shelf life of 21 to 35 days, storage lesions and the potential for blood wastage remain. Storing WB in an additive solution (AS) containing apoptotic inhibitors may help preserve blood cell viability and improve blood quality over extended cold storage.

Study Design and Methods

Non-leukoreduced WB was obtained from healthy individuals and dosed with: AS, AS+Necrostatin-1 (AS+N1), AS+Boc-D-fmk (AS+B; apoptosis inhibitor), AS+Q-VD-OPh (AS+Q; apoptosis inhibitor), and Control (0.9% saline). Blood bags were kept refrigerated (1°–6°C) for 21 days. Bags were tested on days 0, 7, 14, and 21 for complete blood count, metabolism, clot formation, aggregation function, platelet activation, and red blood cell quality.

Results

Platelet count was better preserved in all AS-containing samples. All groups displayed increased glucose consumption and lactate production with storage. Furthermore, all groups displayed a similar decline in clot strength (max amplitude) over the 21-day storage period. Bags that received AS displayed greater preservation of GPIIb expression and lower phosphatidylserine exposure. P-selectin expression was increased in all AS groups.

Discussion

Treatment of hemorrhagic shock with WB transfusion is logistically simpler than component therapy. Results from our study suggest that refrigerated WB stored with an AS containing apoptotic and necrotic inhibitors helps better preserve platelet count but does not improve platelet function. The future development of WB ASs is warranted to optimize both platelet quality and hemostatic function.     

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.     

Effects of prolonged room temperature holding of whole blood intended for preparation of components

Current FDA regulations require that platelet concentrates must be prepared within four hours of whole blood collection. To determine if this time period was critical for the harvesting of viable platelets and other blood components, whole blood was held at room temperature for four, six or eight hours after which various components were prepared. Our results indicate that up to eight hours storage had no detrimental effect on platelet yield, recovery in vivo or lifespan. AHF activity, supernatant hemoglobin and red blood cell ATP were also unaffected.     

Ambient overnight hold of whole blood prior to the manufacture of blood components

Blood services routinely separate whole blood into components that are then stored under different conditions. The storage conditions used for whole blood prior to separation must therefore be a compromise between the needs of the red cells (which benefit from refrigeration) and plasma and platelets (which are better preserved at ambient temperature). For many years, the approach has been to manufacture plasma and platelet components on the day of blood collection, and to refrigerate any unprocessed blood for manufacture into red cell components on the following day. However, this can make it challenging to maintain adequate stocks of all components. The European practice of ‘ambient hold’ of whole blood for up to 24 hours prior to processing allows greater flexibility in blood component manufacture, and the data reviewed suggest there is relatively little impact on the quality of red cell or plasma components, and an improvement in the quality of platelet components.     

Alteration of red cell aggregability and shape during blood storage

BACKGROUND: Storage of blood units (for 35-42 days, depending on the preservative solution) has been reported to induce changes (e.g., reduction of sialic acid level) in red cells that are expected to alter their aggregability. STUDY DESIGN AND METHODS: The aggregability of stored red cells was monitored in their autologous plasma and compared to that obtained with washed cells in dextran-containing buffer throughout the storage period. Red cell aggregability was determined by using a computerized image analyzer of cell flow properties. RESULTS: Blood storage induced changes in red cells that are associated with continuous increase of their aggregability. At the same time, blood storage was associated with a reduction in the level of plasma fibrinogen, the major aggregating agent in plasma. Accordingly, the increased red cell aggregability was observed in red cells stored in dextran-containing buffer, but not in red cells stored in autologous plasma. CONCLUSION: Because blood transfusion is routinely given to patients with normal or high fibrinogen level, the transfusion of stored red cells has the potential to induce increased aggregation in vivo, depending on the storage period. This should be taken into account when blood transfusion is considered, particularly for patients with microcirculatory disorders.