Preparation and storage of platelet concentrates

A technique of platelet concentrate preparation and storage is presented which permits the maximum number of viable and functional platelets to be preserved for periods of 72 hours. Although the storage conditions must be followed precisely, the method is nevertheless simple to perform and does not require specialized expensive equipment. Critical factors include: 1) preparation of the platelet concentrates with an initial centrifugation of 1000 × g for 9 minutes and a second centrifugation of 3000 × g for 20 minutes (86%+/− 1 platelet yield), 2) a storage bag composed of either Fenwal's PL-146 or McGaw plastic, 3) constant gentle mixing, 4) a 70 ml residual plasma volume, and 5) room temperature storage (22 C +/− 2). In Vivo platelet recovery after 72 hours of storage at room temperature averaged 46 per cent +/− 3 and survival was 7.9 days +/− 0.3 (81% of fresh platelet viability). The function of these platelets as measured by the correlation between bleeding time and platelet count after transfusion of pooled platelets into unimmunized, aplastic thrombocytopenic recipients was as good as that of fresh platelets. Both viability and function of concentrated platelets stored at 4 C are severely compromised.     

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.     

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.     

Reduction of the volume of stored platelet concentrates for use in neonatal patients

Premature infants and neonatal patients who require platelet transfusions may develop circulatory overload when administered a 50-ml unit of platelet concentrate. We evaluated the influence of centrifugation and resuspension steps used to reduce the volume of stored platelet concentrates on platelet properties by in vitro methods and by determining post-transfusion increments in neonatal patients. In vitro studies were conducted with platelet concentrates stored at 20 to 24 degrees C for 1 and 5 days in CLX (Cutter) and PL732 (Fenwal) containers and for 1 and 2 days in PL146 containers (Fenwal). With platelets stored in any of the three containers, platelet morphology, mean platelet volume, hypotonic stress response, synergistic aggregation, and platelet factor 3 activity were not affected by the processing steps. The centrifugation and resuspension steps did not cause an enhanced discharge of lactate dehydrogenase from platelets. Similar results were obtained when the platelet concentrates were stored on either a flatbed or an end-over-end tumbler agitator. The in vitro platelet recovery following volume reduction was at least 85 percent. In vivo studies were conducted with platelets stored in the PL732 and PL146 containers. Infusion of platelet concentrates after volume reduction produced a mean corrected increment of 18,947 +/− 14,824 when platelets were stored in the PL146 container and 16,178 +/− 15,699 when platelets were stored in the PL732 container. These results indicate that the volume of stored platelet concentrates can be reduced in a manner which maintains platelet properties.     

Factor VIII preservation following collection into commercial plasmapheresis systems

The two-stage method of Pool and Robinson was used in determining plasma Factor VIII activity following contact with four commercially available plastic blood and pooling bag films. Employing a two-arm double plasmapheresis technique, it was shown that Factor VIII activity in whole blood initially drawn into BB-69 and PL-130 blood bags decreased only slightly from in vivo levels for activity in PL-130 blood bags (t equal 3.2317, p. less than 0.01). Factor VIII activity fell to 98 and 90 per cent preservation levels from an initial 104 per cent in vivo activity, for BB-69 and PL-130 respectively. Factor VIII activity in plasma collected in Ellay and PL-146 pooling bags, both decreased significantly to 85 and 86 per cent, respectively, after three hours storage at room temperature (24 to 25.5 C). Following five weeks storage (-30 C) in Ellay and PL-146 pooling bags, Factor VIII activity significantly (p less than 0.005) fell to 79 and 77 per cent, respectively. No significant differences were observed when Factor VIII activity levels were compared in BB-69 versus PL-130 or Ellay versus PL-146 plastic film systems.     

Evaluation of apheresis platelet concentrates stored for 5 days in PL 732 bags

To assess the functional viability of platelets collected by standard apheresis techniques using the Fenwal CS-3000 "closed system" and stored in Fenwal PL 732 plastic bags for 5 days at room temperature with agitation, a number of in vitro parameters (pH, morphology, platelet volume distribution, osmotic recovery, aggregation, and platelet-associated IgG) were examined as a function of storage time. During the first 24 hours of storage, minimal changes were observed in the test parameters with the exception of ADP-induced aggregation (75% decrease [10 uM], 84% decrease [5 microM]). Significant differences were observed between fresh (day 0) and 5-day-old platelet concentrates in all parameters except median platelet volume. These observed changes in in vitro test parameters with storage time are similar to those previously observed for comparably stored random single-donor platelet concentrates. Thus, the "closed-system" PL 732 apheresis platelet concentrates would be expected to be as effective in vivo as random single-donor platelet concentrates, while minimizing recipient exposure to transmissible agents of infectious disease.     

Applications of ultraviolet light in the preparation of platelet concentrates

Passenger lymphocytes in platelet concentrates (PCs) may induce the formation of lymphocytotoxic antibodies (LCTAbs) and subsequent refractoriness to platelet transfusions. Ultraviolet (UV) irradiation can prevent lymphocytes' acting as stimulator or responder cells in mixed-lymphocyte reactions (MLRs) and could theoretically prevent LCTAb formation in vivo. A system has been devised for the delivery of UV irradiation to PCs; platelet storage characteristics and MLRs were evaluated in UV-irradiated PCs harvested from healthy donors with the Haemonetics V50 and PCS cell separators. MLR and response to phytohemagglutinin stimulation were abolished by a dose of 3000 joules per m2 at a mean wavelength of 310 nm. Platelet aggregatory responses to adenosine diphosphate (ADP), ristocetin, collagen and epinephrine, hypotonic shock response, and pH showed no important differences when control PCs and PCs irradiated as above were compared during 5 days of storage in Fenwal PL-1240 packs. Lactate production during storage was significantly higher in UV-treated PCs (p less than 0.001), but values did not exceed 20 mmol per L. UV transmission at 310 nm in standard blood product containers, including the Fenwal PL-146, PL-1240, and PL-732, was low (less than 30%), but it was acceptable in the Delmed Cryostorage and DuPont SteriCell packs (greater than 50%). UV irradiation may provide a simple and inexpensive means of producing nonimmunogenic PCs.     

Di-2-ethylhexyl phthalate (DEHP) and mono-2-ethylexyl phthalate (MEHP) accumulation in whole blood and red cell concentrates

Plasma DEHP concentrations were measured weekly in whole blood and red cell concentrates (RCC) during 21 days of storage in standard CPD within PL-130 blood bags. In addition, DEHP and MEHP accumulation patterns were investigated in blood stored for 42 days in modified CPD with adenine within PL-146 and BB-69 storage containers. Total per-unit plasma DEHP of RCC units was 49 to 71 per cent of the total in plasma of whole blood units (PL-130). From 28 to 42 days, mean DEHP levels were 12 to 19 per cent higher in whole blood stored in PL-146 than in BB-69. Although MEHP was not found in any blood bag plastic, MEHP accumulated in plasma during whole blood storage. MEHP concentrations were 2.8 to 3.8 times higher in plasma stored in BB-69 than in PL-146. It is postulated that MEHP arises from hydrolysis of DEHP by plasma lipase, even in frozen plasma sample, and that the rate of this reaction is influenced by blood bag plastic surface characteristics.     

Growth of gram-positive and gram-negative bacteria in platelet concentrates

In 1986 the allowable platelet storage time was reduced from 7 to 5 days because of a recent increase in septic deaths associated with platelet transfusion. In this study, the growth curves of two gram-positive and two gram-negative organisms in platelets stored for 7 days in CLX and PL-732 bags were evaluated. Platelets in CLX bags were inoculated with 10(1), 10(2), and 10(3) organisms and 10(2) organisms were introduced into PL-732 bags. Test organisms were inoculated into trypticase soy broth as a control. All four bacteria grew rapidly in trypticase soy broth, reaching 10(9) organisms per mL within 48 hours. In both CLX and PL-732 bags, the growth pattern of gram-positive organisms was generally logarithmic during the first few days of storage. A concentration of 10(8) organisms per mL was present by Day 3 or 4, after which further proliferation was inhibited by the high density of bacteria in the platelets. In PL-732 bags, the proliferation of gram-negative organisms followed a pattern similar to that of the gram-positive bacteria. However, gram-negative organisms grew less well in CLX bags.(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)     

Extended storage of single-donor platelet concentrate collected by a blood cell separator

Use of a sealless blood pathway in a blood cell separator (CS-3000, Fenwal) permits collection of platelets in a "closed system" when saline and anticoagulant solutions are integrally attached; this in turn allows storage of instrument-collected platelet concentrates (PCs) beyond 24 hours. To evaluate extended storage of high yield PCs, cells collected with the instrument were stored (200 ml plasma) for 8 days (flatbed agitation) in either 3-liter polyvinylchloride (PL 146) containers (n = 6), polyolefin bags (PL 732) (n = 8), or two 1-liter polyolefin (double PL 732) containers (n = 8). A mean of 4.45, 4.09, and 3.94 X 10(11) platelets were stored in PL 146, single PL 732, and double PL 732, respectively; total white cells per container averaged 0.3, 0.2, and 0.2 X 10(9) for the three container systems. By day 1, platelet pO2 dropped to 14 and 16 torr in PL 146 and PL 732 PCs (pCO2, 127, and 82 torr). In contrast, double PL 732 maintained high pO2 (approximately equal to 80 torr) and low pCO2 (approximately equal to 30 torr) through day eight. Glucose declined at faster rates in PL 146 and single PL 732 containers, while lactate increased more rapidly (338 and 197 mg/dl of lactate on day four vs. 116 mg/dl for double PL 732 units). Morphology scores dropped from 400 to 98 (PL 146) and 216 (PL 732) at day four (pH values of 6.3 and 7.0), while a score of 330 was seen in double PL 732 PCs.(ABSTRACT TRUNCATED AT 250 WORDS)     

Extended storage of platelets in a new plastic container. II. In vivo response to infusion of platelets stored for 5 days

A new blood container material (PL 1240 plastic) made of polyvinyl chloride containing a tri(2-ethylhexyl) trimellitate plasticizer was evaluated in three laboratories. When platelet concentrates (50-60 ml) were stored on a variety of agitators for 7 days at 22 +/- 2 degrees C, poststorage pH (mean +/- SD) ranged from 7.29 +/- 0.05 (6 rpm elliptical rotator) to 6.87 +/- 0.8 (70 cycles per minute flatbed agitator). The platelet counts ranged from 1.51 +/- 0.12 to 0.95 +/- 0.36 X 10(6) per microliter. Morphology scores and hypotonic shock response values of platelets stored 7 days in PL 1240 plastic containers were better than those noted following 3-day storage of control platelets in PL 146 plastic containers. The percent discharge of lactic dehydrogenase from platelets stored 7 days in PL 1240 plastic containers for 3 days (p less than 0.05). Mean platelet recoveries of 44 +/- 15 percent (n = 11; 111Indium) and 39 +/- 8 percent (n = 29; 51Chromium) were seen when autologous platelets were infused following 5-day storage in PL 1240 plastic bags. Platelet half-lives of 3.6 +/- 0.4 (n = 9) 4.1 +/- 0.4 (n = 20) days were reported in the two laboratories which used 51Cr labeling, while survival values of 7.0 +/- 1.0, 2.8 +/- 0.8, and 5.4 +/- 1.9 days were seen when data from the 111Indium studies (n = 11) were analyzed using linear, exponential, and multiple hit programs, respectively. Platelets stored for 5 days also were administered to 13 thrombocytopenic oncology patients.(ABSTRACT TRUNCATED AT 250 WORDS)     

Paired comparison of the in vivo and in vitro results of storage of platelet concentrates in two containers

Both in vitro and in vivo methods are used to test the validity of techniques for storing platelet concentrates for transfusion. In this study, the characteristics of platelet concentrates stored for 5 days at 22 degrees C in two different containers were evaluated by paired comparison using two in vitro measurements and two in vivo measurements. On two occasions, 10 normal subjects donated concentrates that were stored in containers of either the CLX system or the PL-146 system. The first plastic used was chosen at random. If necessary, a concentrate platelet count was reduced to 1,200,000 per microliters by addition of plasma to avoid pH fall. Mean recoveries were 48.2 +/- 10.6 percent (mean +/- 1 SD) and 42.4 +/- 7.8 percent for platelets stored in containers of the CLX and PL-146 systems, respectively. Similarly, survivals (T 1/2 in days) were 3.4 +/- 0.8 and 3.0 +/- 0.7, respectively. Since a paired design was used, the superiority of the CLX system was demonstrable with a one-tailed paired t test. If a paired design had not been used, a pooled t test would have been appropriate and the differences would not have been significant. This result emphasizes the value of the paired design. Furthermore, two in vitro measurements that reflect platelet morphology, dispersion of the size distribution and extent of shape change with adenosine diphosphate, were superior for platelets stored in CLX containers as well, suggesting a relationship between these measurements and in vivo viability.(ABSTRACT TRUNCATED AT 250 WORDS)     

5-day storage of platelets collected on a blood cell separator

One of the earliest devices available for plateletpheresis is the Haemonetics system; this machine has been updated recently to permit the use of software in a closed system and thus storage of the collected platelets beyond 24 hours. The authors examined the in vitro and in vivo function of platelets collected on the Haemonetics AutoSurge machine and stored for 5 days in two separate 1000-mL CLX bags. The average count per bag was 1.8 +/- 0.4 x 10(11) in approximately 200 mL of plasma. Immediately following collection, the platelet response to ADP and epinephrine represented 78 and 35 percent, respectively, of the preapheresis values. Aggregation to single stimuli subsequently decreased to 29 and 0 percent, respectively by Day 5. This response is equal to or better than the response reported with the Fenwal CS-3000, the only other plateletpheresis device routinely used for long-term storage. The pH of the preparations was well maintained throughout storage, and there was little alteration in hypotonic shock response or serotonin uptake. Serotonin release decreased consistently. The morphology scores indicated good maintenance of shape immediately following collection; this subsequently decreased after 5 days of storage. Bacterial cultures were negative in all instances. The 51Cr in vivo survival and recovery was good with 65.5 +/- 7.1 percent recovery and an average survival of 7.3 +/- 1.3 days (multiple hit; n = 5). The data indicate that storage of the Haemonetics plateletpheresis product is feasible and that the product is as good as others currently available.     

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.     

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.     

Comparative Studies of the Viability of Human Platelets Stored at 4 C in Plastic and Glass Containers

Studies were done to investigate whether the surface of the container (plastic or glass) would influence the viability of human platelets stored at 4C for short intervals.
Cr⁵¹-labeled platelets prepared as concentrates suspended in plasma were preserved at 4 C for 24, 48 or 72 hours and their capacity to recirculate and survive after infusion into the respective donors ("viability") was determined. In addition, the clot retraction property of the stored platelets was measured.
Platelet viability was sharply reduced after storage at 4 C in all experiments. When the storage period was limited to 24 hours, survival curves of platelets preserved in plastic bags were similar to those of platelets preserved in glass bottles. However, after 48 hours of storage, viability of platelets preserved in plastic containers had values significantly higher than those of platelets preserved in glass containers. After 72 hours of storage, platelet viability was reduced to minimal values but was still greater for platelets stored in plastic bags.
The study of clot retraction confirmed data previously obtained and showed that this platelet property was preserved better by storage in glass bottles rather than in plastic bags. Platelet viability was, however, lost very rapidly during storage at 4 C in either type of container so that the unfavorable effect of plastic on the preservation of clot retraction was thought not to be of practical importance in platelet transfusion therapy.     

Serotonin uptake at 22 degrees C of stored platelets

In order to ascertain the possibility that platelet serotonin uptake may occur during storage of platelet concentrates (PC) at 22 degrees C with agitation, the high-performance liquid chromatographic procedure was applied to determine serotonin uptake by platelets. Studies at 22 degrees C showed that platelets stored for 4 days exhibited a significant serotonin uptake with a Vmax value of 2.4 X 10(-19) mole/platelet/min and a Km value of 0.62 X 10(-6) M. Incubation of PC with 5 X 10(-6) M serotonin for 1 day at 22 degrees C increased their serotonin contents from 2.2 to 4.2 X 10(-7) mole/10(11) platelets. Thrombin stimulation caused about 80% release of intracellular serotonin from fresh as well as stored platelets, which contained standard serotonin in the same amount as the original amount. These results suggest that a significant serotonin uptake of platelets might occur during in vitro storage at 22 degrees C and stored platelets have retained abilities to sequester extracellular serotonin into dense granules.     

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.     

Recovery, lifespan, and function of CPD-Adenine (CPDA-1) platelet concentrates stored for up to 72 hours at 4 C

We studied the in vivo recovery, lifespan, and hemostatic effectiveness of CPDA-1 platelet concentrates stored for up to 72 hours at 4 C. A total of 120 CPDA-1 concentrates containing an average (+/− 1 S.D.) of 6.6 +/− 2.0 × 10(10) platelets were prepared. The pH of the units following storage at 4 C was 6.8 +/− 0.2; no unit had a pH below 6.3. Autologous transfusion of six normal volunteers showed that platelets stored at 4 C for 72 hours had an in vivo recovery of 40 +/− 18 per cent and a lifespan of 5.1 +/− 1.5 days. The hemostatic effectiveness of CPDA-1 platelets was determined by platelet counts and template bleeding time measurements in 10 thrombocytopenic patients. Patients receiving 48-hour-stored platelets had a four- to six-hour posttranfusion corrected platelet increment averaging 15,300 +/− 3,200/microliter which was 67 +/− 34 per cent of expected recovery. Four of the five patients transfused with this preparation showed an improved bleeding time. In contrast, three patients receiving 72-hour- stored platelets had a four- to six-hour posttransfusion increment of 5,800 +/− 2,400/microliter that was only 26 +/− 13 per cent of the expected recovery; furthermore, only one of these patients showed any correction of the bleeding time. These data indicate that CPDA-1 platelets are hemostatically effective when stored at 4 C for up to 48 hours.