Platelet storage for 7 days in second-generation blood bags

Plastic storage bags designed to optimize O2 and CO2 transfer to preserve platelets for 7 days prior to transfusion were studied in vivo and in vitro. Platelets stored 7 days in second-generation CLX bags were compared to platelets stored 3 days in standard (CL-3861) 3-day storage bags and platelets transfused within 24 hours of collection. The CLX bags maintained concentrate pH at a mean of 6.85 +/- 0.03 (SEM) after 7 days, while in standard bags after 3 days of storage, the mean pH was 6.46 +/- 0.03. A smaller proportion of platelets stored 7 days in CLX bags were discarded because of a pH less than 6.0 compared to those stored 3 days in CL-3861 bags (10 vs 21%). Poststorage pH showed strong correlation with concentrate platelet count and weak correlation with concentrate white cell count in both bag types. There was no significant difference in the mean corrected platelet count increments between platelets stored 7 days in second generation CLX bags and those stored 3 days in CL-3861 bags (10,000 and 12,200 at 1 hour, and 7000 and 7500 at 24 hours, respectively) following transfusion to 16 thrombocytopenic recipients. However, transfusion of fresh platelets achieved mean corrected increments at both 1 and 24 hours posttransfusion that were higher than seen with either group of stored platelets (20,100 at 1 hour and 10,800 at 24 hours). Platelets can be stored 7 days in second-generation CLX blood bags with results comparable to those of platelets stored 3 days in standard bags.     

Evaluation of a new, more oxygen-permeable, polyvinylchloride container

A new container made of polyvinylchloride (PVC) with diethylhexyl phthalate (DEHP) used as the plasticizer was subjected to in vitro and in vivo evaluation for prolonged platelet storage. As compared with the original PVC bag, this bag has increased gas permeability by its reduced film thickness, larger surface area (400 ml capacity), and more porous label. The oxygen permeability coefficient, K(O2), of the new container was measured to be 655 nmol per min per atm. On the basis of previous studies relating the K(O2) to the maximal platelet count, it was predicted that this maximal count would be in the range of 7.9 to 8.9 X 10(10) platelets. This prediction was confirmed by carrying out 58 studies measuring pH, pO2, and platelet count on platelet concentrates (PCs) stored for up to 7 days. After 5 days of storage all PCs with counts above 8.0 x 10(10) had pH less than or equal to 7.0, whereas those with counts below 8.0 x 10(10) had pH greater than or equal to 7.0. Six units (10%) with counts above 9.0 X 10(10) had pH levels of 6.5 or below. Thirteen of the PCs underwent extensive in vitro testing of platelet function during 7 days of storage. No significant differences were found in pH, ATP content, and decrease in platelet count, as compared with studies (n = 22) using PCs stored in polyolefin containers.(ABSTRACT TRUNCATED AT 250 WORDS)     

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.     

Seven-day storage of apheresis platelets: report of an in vitro study

BACKGROUND: The objective of this study was to determine the allowable platelet content limits for apheresis platelets stored for 7 days in a platelet storage bag (COBE ELP, Gambro BCT). METHODS: Apheresis platelets under controlled concentration and volume per bag were stored in plasma up to 8 days at 22 degrees C with horizontal agitation. Routinely evaluated in vitro platelet parameters were followed. Oxygen consumption was directly measured with a Clark-type electrode. All components were cultured in aerobic medium on Day 7. RESULTS: Twenty-four components were evaluated in storage configurations (median [range], 340 [110-402] mL, 1.32 [0.99-2.45] x 10(6) platelets/microL, and 4.8 [1.4-5.9] x 10(11) platelets/bag). No bacterial contamination was detected. One component had a pH value at 22 degrees C of below 6.0 before Day 5 with attendant loss of all other in vitro function measures. The pH value at 22 degrees C was maintained above 6.2 for the remaining 23 components. A pH value of greater than 7.4 was observed at some point in storage for 13 of 23 units, although platelet function or activation was not adversely affected. Aerobic metabolic function was maintained over 7 days with O2 consumption of 321 micromol per hour per 10(12) platelets on Day 7. CONCLUSION: Although a continuing decline of platelet in vitro characteristics can be observed for storage beyond 5 days, apheresis platelets in plasma stored 100 to 400 mL per bag, 1.0 x 10(6) to 2.5 x 10(6) platelets per microL, and a maximum of 5.1 x 10(11) platelets per bag maintained in vitro platelet characteristics over 7 days of storage.     

Extended storage of platelets in a new plastic container. I. Biochemical and morphologic changes

A new polyvinyl chloride container plasticized with tri(2-ethylhexyl) trimellitate (PL 1240 plastic) was evaluated for use in extended platelet storage. Six leukocyte-rich platelet concentrates (mean, 0.6 X 10(9) white cells per bag; range, 0.3 to 1.0 X 10(9) per container) were prepared by removing as much of the platelet-rich plasma from blood as possible. The cells were stored at 22 degrees C on an end-over-end agitator. An average of 1.04 +/- 0.19 X 10(11) platelets was recovered, and the mean pH dropped from 7.23 on day 0 to 6.68 by day 5. At the completion of the storage period. PO2 averaged 80 torr, PCO2 was 35 torr, bicarbonate concentration was 0.5 mM, and lactate concentration 29.5 mM. Thirty-one additional units of platelet concentrates, not deliberately prepared to be leukocyte-rich, on day 5 had a pH of 6.75 +/- 0.39 (mean platelet yield, 0.97 +/- 0.21 X 10(11); PO2 and PCO2 averaged 50 and 48 torr, respectively). Following storage, the cells had an average phase microscopic morphology score of 244 (n = 17). Platelets appeared to be preserved well throughout storage when assessed by transmission and scanning electron microscopy. We conclude that platelets can be stored for 5 days in PL 1240 plastic containers with good preservation of pH and cell ultrastructure.     

Effect of white cells on platelets during storage

White cells (WBCs) present in stored platelet concentrates have adverse effects on platelet function and on posttransfusion recovery. Although these effects have been attributed to the fall in pH that results from active WBC metabolism, platelets stored in gas-permeable storage bags still exhibit abnormalities, despite maintenance of a stable pH of greater than 6.0. The changes in platelet proteins and function brought about by storage with a controlled number of WBCs were studied. Twelve platelet-pheresis specimens were centrifuged at 180 x g to achieve a WBC count of less than 2 x 10(5) per mL (which contained less than 10% granulocytes). These specimens were split into two aliquots and placed in platelet bags for storage at 22 degrees C with constant horizontal agitation. Neutrophils, obtained from the same donor by centrifugation of 50 mL of whole blood through a discontinuous ficoll gradient, were added to one of the two platelet storage bags to achieve a final neutrophil count of 1 x 10(6) per mL. Platelet aliquots were removed and studied on Days 3 and 5. In platelets stored without neutrophils, the average response to ristocetin, using the mean slope as an index of platelet responsiveness, was 10.3 (n = 9, SD = 11) on Day 3, whereas for the platelets stored with neutrophils, it was 1.25 (n = 12, SD = 0.9, p less than 0.01). Significant differences were also seen on Day 5 (slope = 4.5 for platelets stored without neutrophils, slope = 0.3 for platelets stored with neutrophils, p less than 0.01). Platelet aggregation with 8 microM ADP and 1.5 mg per mL of collagen did not differ significantly.(ABSTRACT TRUNCATED AT 250 WORDS)     

Five-day storage of platelet concentrates

The short 72-hour shelf-life of platelet concentrates stored in standard PL146 (Fenwal) plastic bags often results in shortages of platelets. This 3-day limitation is based on the biochemical and physiological changes that occur during storage and that result in decreased viability and survival after transfusion. We assessed both in vitro and in vivo function of platelet concentrates stored for 3 and 5 days in two new plastic packs: PL732 (Fenwal) and CLX (Cutter). The concentrate pH was maintained above 7.0 in both bags and there was little change in platelet count or size following 5 days of storage. Aggregation response to adenosine diphosphate, epinephrine, and collagen was maintained well. The PCO2 values indicated good gas escape with lower values after 5 days of storage than at 0 time. Lactate accumulation and glucose utilization were also lower in these new bags. Autologous survivals of chromium-labeled platelets stored for 5 days were 6.0 days (PL732) and 5.1 days (CLX), which are equal to or better than those found for platelets stored for 3 days in PL146. Posttransfusion increments in thrombocytopenic patients were acceptable; 49 percent after 1 hour and 31 percent after 24 hours for concentrates stored in CLX and 44 percent after 1 hour and 28 percent after 24 hours for concentrates stored in PL732. Both of these new bags, which contain different types of plasticizers, provide an environment that results in an improved product and will permit 5-day storage of platelet concentrates; these two benefits will help to alleviate the difficulties in supply of platelet concentrates.     

Extension of platelet concentrate storage

Extension of the storage time of platelet concentrates in a satellite bag which is part of a new blood bag system was studied by reinfusing autologous 51Cr-labeled platelets into normal volunteers, and measuring postinfusion platelet counts and bleeding times in patients requiring platelet transfusions. This satellite bag, made of polyvinylchloride plasticized with a new agent, was found to protect platelet concentrates against fall of pH better than other containers studied. This protection was felt to be due to the greater gas permeability of the new plastic. Mean in vivo recovery and half-life (greater than 31% and 3.3 days, respectively) of autologous reinfused platelets were satisfactory following 5 days of storage. Following 7 days of storage, mean recovery was 41 percent and half-life was 2.8 days. Peripheral platelet count increments in patients following platelet transfusions with concentrates stored 4 to 7 days in the new plastic were comparable to increments following transfusion of platelets stored 2 to 3 days in the other plastics studied. Bleeding times shortened in three of four patients receiving platelet concentrates stored from 4 to 6 days in the new plastic. Platelet concentrates stored in the new bag at 20 to 24 degrees C with flat-bed or elliptical agitation could be transfused for up to 5 days following phlebotomy with acceptable clinical results. The new plastic container is promising for storage of platelet concentrates for up to 7 days. Due to the higher pH of 50-ml platelet concentrates stored in bags made with the new plastic, the concentrates were superior at any storage interval to those stored in bags made of the other plastics studied.     

Storage of single donor platelet concentrates: Paired comparison of storage as single or double concentrates

Modern cell separators allow the collection of two plateletpheresis concentrates (PCs) at one session. This study evaluates the quality of PCs stored as double concentrates in standard storage containers of two manufacturers. We collected 20 PCs that contained 4.5 × 10¹¹ platelets in 375 ml plasma (10 using the COBE Spectra and 10 using the Fresenius AS.TEC 204 with 500 ml bags) that were split into one unit of 3.0 × 10¹¹ platelets in 250 ml (3.0‐PC) and one of 1.5 × 10¹¹ platelets in 125 ml (1.5‐PC). Storage of one 3.0‐PC per bag of a two‐bag system corresponded to storage conditions for double PCs and storage of one 1.5‐PC per bag to storage conditions of single PCs. Cell counts, blood gas analysis, glucose and lactate levels, platelet aggregation, and activation and plasma levels of β‐ thromboglobulin (β‐TG) and complement factor 3a (C3a) were measured before storage and again on days 3 and 5. COBE 3.0‐PCs demonstrated less pH rise, lactate production, CD 62P expression and β‐TG plasma levels, and better aggregability after storage than COBE 1.5‐PCs. Fresenius 1.5‐PCs had similar platelet quality to COBE 3.0‐PCs. Fresenius 3.0‐PCs showed a fall of pH (day 5: 6.22 ± 0.56), the highest amount of anaerobic glycolysis compared to all other storage conditions investigated, high CD 62P‐ expression and β‐TG plasma levels, and impaired aggregability on days 3 and 5. The highest C3a levels were found in COBE 1.5‐PCs. 3.0 × 10¹¹ platelets in 250 ml plasma should be stored either in one bag of the COBE system or in two 500 ml bags of the Fresenius system. The COBE two‐bag system allows the storage of two PCs without loss of platelet quality. Two PCs should not be stored in the Fresenius C4L 500 ml storage containers. J. Clin. Apheresis. 16:148‐154, 2000. © 2001 Wiley‐Liss, Inc.     

Effective leukocyte removal from platelet preparations by centrifugation in a new pooling bag

A pooling bag (Leukotrap, Cutter Laboratories, Berkeley, CA) with a protruding pocket at the bottom into which heavier cellular elements are collected after centrifugation at 390 X g for 10 minutes, was studied. The tab is clamped before transfusion. Varying numbers of platelet concentrates (PC) that had been stored for different durations were pooled and centrifuged in the bags. When 4 or more units of PC were studied, the results were independent of the number of units and the duration of storage. Approximately 90 percent of the contaminating leukocytes (WBC) were removed with a platelet loss of less than 10 percent. Similar results were obtained with single-donor platelets (WBCs decreased 93%; range, 77-99%; n = 12). Posttransfusion increments were similar to those with unmodified platelets; in four patients, febrile platelet transfusion reactions were eliminated by the WBC removal. Thus, the bags represent a simple, reproducible, and effective means of reducing WBC and red cell contamination of platelet preparations with acceptable platelet losses.     

The effect of mode of agitation and type of plastic bag on storage characteristics and in vivo kinetics of platelet concentrates

We studied the characteristics of platelet concentrates stored for 5 days at 22 degrees C. Platelets were prepared in three plastic bags (PL 732, PL 1240, and CLX) and stored on one of four platelet agitators, 1- or 6-rpm elliptical and 2- or 6-rpm circular rotators. A total of 76 studies were divided among 12 groups, each group being composed of a different storage bag-rotator combination. In vivo recovery and survival were calculated using Indium-111 oxine-labeled platelets injected into autologous volunteers. Platelet recovery was assessed at 2 hours postinjection or as the y-intercept of the multiple-hit model survival curve. Survival was calculated using linear, exponential, and multiple-hit computer models. Linear T 1/2 also was calculated as an index of platelet survival. At 5 days, the pH of all concentrates was above pH 7.0 and platelet counts were above 5.5 X 10(10) per bag except for the PL 732 with the 6-rpm elliptical rotator, which was 4.6 X 10(10) per bag. This combination also showed a significantly higher poststorage lactic dehydrogenase (LDH) discharge compared to the mean of the other 11 groups (23.6 +/- 5.4% vs. 10.4 +/- 3.0%, p less than 0.05); however, the beta-thromboglobulin (beta-TG) release was not statistically different.(ABSTRACT TRUNCATED AT 250 WORDS)     

Extension of platelet concentrate storage to 7 days in second- generation bags

Platelet concentrates stored for 7 days in 50 ml of plasma in both thin film and enlarged variations of the standard 5-day CLX plastic bags were evaluated for pH maintenance and in vivo viability by two laboratories working independently. 51Cr-labeled platelets were reinfused into normal volunteers at the end of storage and recovery and half-life calculated. The pH was maintained well; less than 10 percent of units fell below 6.0 at 7 days. Mean 7-day recovery for both laboratories was 43.6 +/- 11.6 percent in the thin-film bag and 45.4 +/- 8.52 percent in the enlarged bag, compared with 43.6 +/- 8.8 percent at 5 days in the 5-day plastic licensed bag. After 7 days storage the half-life was 3.6 +/- 0.9 days in the thin-film bag and 3.7 +/- 0.6 days in the enlarged bag, compared with 3.6 +/- 0.5 days in the previously licensed CLX plastic bag after 5 days. Thus, platelet viability was maintained well at 7 days of storage in both of the container variations that allowed increased gas exchange.     

Platelet storage in Fresenius/NPBI polyolefin and BTHC-PVC bags: a direct comparison

SUMMARY: New platelet storage systems, such as changes in the plastic of the storage bags, require validation. In this study, pooled buffy coat platelets stored in Fresenius/NPBI polyolefin bags were compared with those stored in Fresenius/NPBI butyryl-trihexyl citrate (BTHC) plasticized polyvinyl chloride (PVC). The CompoSelect thrombocyte polishing filter system (1000 mL polyolefin bag) and the CompoStop F730 system (1300 mL BTHC-PVC bag) were used to prepare paired, plasma-suspended, buffy coat platelet concentrates. Samples were taken up to day 7 for in vitro analysis. In a separate experiment, 12 units were prepared using the CompoStop F730 system and samples taken after leucofiltration for FXIIa assay. By day 7, platelet concentrates stored in BTHC-PVC demonstrated significantly higher pH levels (7.32 +/- 0.05 vs. 7.26 +/- 0.05) and a greater degree of cell lysis as shown by increased lactate dehydrogenase levels (497 +/- 107 vs. 392 +/- 81 U L(-1)). The supernatants contained higher concentrations of soluble P-selectin and the chemokine 'regulated on activation, normal T-cell expressed and presumably secreted', which are released from the alpha-granules during activation. The ATP concentrations were significantly lower in BTHC-PVC. Platelet counts, mean platelet volume and hypotonic shock response were similar for both bags. FXIIa antigen concentrations were 0.6 +/- 0.2 ng mL(-1) indicating that activation of the contact factor pathway had not occurred. Although the CompoStop F730 leucoreduction filter did not activate the contact system, platelets stored in 100% plasma in BTHC-PVC bags demonstrated different in vitro characteristics from those stored in polyolefin. Further work is required to demonstrate whether these differences will affect in vivo recovery and survival.     

Concentration of platelet units into small volumes

To determine the best procedure for concentrating platelets in a smaller volume after storage, we studied platelet loss after concentration at various centrifugation g forces for various times. In vitro studies demonstrated the need to let platelets sit for 1 hour prior to resuspension by gentle kneading. 500 X g gave unacceptable results. Losses were minimal at 1500 X g for 7 minutes, 2000 X g for 10 minutes and 5000 X g for 6 minutes. Reinfusion of 51 Cr-labeled platelets into normal volunteers after concentration at both 2000 X g for 10 minutes and 5000 X g for 6 minutes showed normal viability. Numbers and viability of platelets stored up to 5 days in 50 ml plasma and then concentrated in 10 ml plasma after centrifugation at 1500 X g for 7 minutes, 2000 X g for 10 minutes or 5000 X g for 6 minutes should be clinically acceptable.     

5-day storage of single-donor platelets obtained using a blood cell separator

Platelets were collected from normal donors via a blood cell separator (Fenwal CS-3000). Platelets were stored initially in two separate 1000-ml bags (average count per bag, 2.3 +/- 0.5 x 10(11)) in 100 ml of autologous plasma for 5 days. Little change in platelet count was noted after 5 days of storage; however, the white cell count fell from 5.1 +/- 1.7 x 10(9) at Time 0 to 3.4 +/- 2.3 x 10(9) per l at Day five. The initial lactate values were 32 +/- 11 mg per dl and rose to 165 +/- 28 mg per/dl by 5 days. Platelet aggregation was impaired both by the collection procedure and during storage: whereas the response to ADP of the donors' platelets before the procedure was 100 percent, samples taken from the product immediately after collection had only a 45 percent response, which fell to 12 percent by Day 5. Aggregation using epinephrine was similarly affected, with a 75 percent response after collection and 0 percent response by Day 5. The plasma beta-thromboglobulin (beta-TG) level was high, both after collection (5.0 micrograms/ml at Time 0) and after storage (11.0 micrograms/ml), indicating a considerable effect of collection on platelet alpha granule release. In vivo recovery of these platelets was very good at 67 +/- 6 percent, with an average survival of 7.3 +/- 1.4 days (multiple hit; n = 4).(ABSTRACT TRUNCATED AT 250 WORDS)     

Storage of pooled platelet concentrates. In vitro and in vivo analysis

The use of sterile connecting devices will permit up to 5-day storage of pooled platelet concentrates (PCs). However, there are no data evaluating long-term storage of PCs pooled from multiple donors. Four units of ABO-compatible or -incompatible PCs were pooled and stored in single 300-ml PL-732 storage bags for up to 5 days. Results of in vitro assays showed acceptable storage values regardless of the ABO types in the pool. Pool pH on Day 5 was 6.83 +/- 0.3 (mean +/- 1 SD). The in vitro storage characteristics were comparable to those of unpooled age-matched platelets reported previously from our laboratory. For in vivo studies, 4-unit pools of ABO-compatible random-donor PCs stored for up to 96 hours in 1000-ml PL-732 bags were transfused into patients who were thrombocytopenic due to bone marrow failure, and the correct count increments (CCI) were determined. In vivo results showed a mean 1-hour CCI of 11,368 +/- 5824 for the pooled stored platelets and 7819 +/- 5189 for unpooled controls (p greater than 0.05). To evaluate the possibility that passenger lymphocytes in the concentrates would generate mixed lymphocyte reactions (MLR) in the pooling bag during storage, lymphocytes were studied over 5 days of storage by the use of monoclonal antibodies against activated T-cell markers and by 3H thymidine uptake. Results failed to show evidence of either the generation of activated T-cell markers or the uptake of 3H thymidine.(ABSTRACT TRUNCATED AT 250 WORDS)     

In vitro characteristics of volume-reduced platelet concentrate stored in syringes

For convenience, small volumes of platelet concentrate (PC) intended for neonatal patients are often dispensed in syringes. The PC, however, may remain in the syringe for up to several hours before the actual transfusion. As there are few data on the effect of such syringe storage on PCs, the in vitro syringe storage properties of small volumes of 1- and 5-day-old units, and volume-reduced units of PC were evaluated. In four separate experiments, PCs were stored in syringes in volumes of 10, 15, or 30 mL for up to 6 hours at 20 to 24 degrees C without agitation. Platelets were evaluated for pH, platelet count, and a variety of biochemical and in vitro functional assays. Results showed that even with the equivalent of a full unit of platelets stored in the syringe for up to 6 hours, the pH did not fall below 6.0. Although there was an increase in lactate production and consumption of glucose, which paralleled the decline in pH, the changes were not greater than those seen in platelets stored up to 5 days in gas-permeable blood bags. Similar results were seen for PCs stored in syringes for 6 hours at 37 degrees C. All of the pH levels recorded at the end of 6 hours of syringe storage were above the minimum required level of pH 6.0. Data from in vitro platelet assays imply that at any time during their shelf life, PCs can be stored in gas-impermeable polypropylene syringes for up to 6 hours and can maintain acceptable storage characteristics; in vivo data are needed to confirm these observations.     

Extension of platelet concentrate storage by addition of sodium bicarbonate

Viability of platelet concentrate (PC) stored in polyvinylchloride bags in an elliptical rotator at 22 degrees C (standard PC) was assessed by in vitro tests, and an alternate approach to extending the shelf-life of PC by the addition of hypertonic sodium bicarbonate (test PC) was investigated. The fall in the pH which occurred during storage in standard PC was arrested in test PC. Furthermore, platelets stored under these test conditions maintained their morphology better than in standard PC as judged by their mean platelet volume and platelet distribution width. Recovery of stored platelets from hypotonic shock at 37 degrees C following resuspension in fresh plasma was better for test platelets. Results indicated that platelets in standard PC were viable up to day 3 but were not viable at day 7. Platelets store better in PC to which sodium bicarbonate has been added and behave as viable platelets up to 7 days.     

Platelet storage. Effects of leachable materials on morphology and function

A polyolefin plastic (PL 732) bag formulated without liquid plasticizer allows storage of platelets for 5 and, now, up to 7 days. In order to assess the leaching of compounds from this new plastic, extracts of the supernatant from platelet concentrates stored in these bags were analyzed by high-performance liquid chromatography, mass spectrometry, and gas-liquid chromatography. A leachable material was detected and identified as di(2-ethylhexyl) phthalate (DEHP). During the sterilization process, migration of the DEHP occurs from the polyvinylchloride (PVC) bags into the PL 732 plastic bag. The level of DEHP was 12-fold less in the extracts of PC supernatant stored in the PL 732 bag than those in the polyvinyl chloride (PL 146) plastic bags which were used previously for platelet storage. Platelets stored in low DEHP concentrations in the PL 732 bags were composed of 10 to 35 percent of unclassifiable shapes. These shape changes were not observed in higher concentrations of plasticizer, although the morphology scores decreased during storage in PL 146 as well. This effect on morphology was not related directly to the dose of DEHP. When platelet membranes were isolated from platelets stored in the presence of radiolabeled DEHP, the amount of bound 14C-DEHP was found to be directly proportional to the concentration of DEHP in the plasma supernatant. However, while there was a linear relationship between the protein concentration in the membrane fraction and the amount of bound DEHP, no specific DEHP binding site could be identified by electrophoresis of the solubilized platelet membranes.     

Collection of platelets with a new cell separator and their storage in a citrate-plasticized container

A new apheresis device using microprocessor control for the collection of a high-purity single-donor platelet concentrate was evaluated, as was the storage of platelets for up to 5 days in a citrate-plasticized polyvinylchloride blood bag. The study was conducted in three phases: collection of platelets for in vitro studies and determination of donor safety; autologous transfusion of platelets in healthy volunteers; and transfusion of platelets in patients requiring platelet transfusion therapy. Donors had mild hypocalcemia and minimal changes in blood counts except for a platelet count reduction from 288 +/- 50 x 10(3) (288 +/- 50 x 10(9)/L) to 217 +/- 43 x 10(3) per microL (217 +/- 43 x 10(9)/L). A mean of 3.36 +/- 1.24 x 10(11) platelets was collected in the mean volume of 214 mL with red cell and white cell contamination in the range of 10(7). Morphology and aggregation were as described previously in stored platelets. Platelet survival data in eight subjects showed a mean recovery of 61 +/- 11 percent and mean survival of 5.03 +/- 1.07 days by a weighted-mean model. Patients transfused with platelets had mean increments of 23,000 immediately and of 8000 at 24 hours; corrected count increments were 6000 at 1 hour and 4000 at 24 hours. The platelets were successful in providing hemostasis to these patients. Clinically useful 5-day-stored platelets are obtained by using this apheresis technology with a functionally closed system and a citrate-plasticized blood bag.