C. R. Valeri

Effect of 24-hour whole-blood storage on plasma clotting factors.

BACKGROUND: The current requirements for the preparation of fresh‐frozen plasma within 8 hours of whole‐blood collection were designed to maintain clotting factor activities. These requirements, however, limit the production of fresh‐frozen plasma in a large blood center. There are few data on the effect of the extension of CPD whole‐blood storage to 24 hours on clotting factor activity.

An Experiment with Glycerol-Frozen Red Blood Cells Stored at –80°C for up to 37 Years

Background and Objectives: Red cells frozen using 40% W/V glycerol are currently FDA approved for frozen storage at –80°C for up to 10 years. Materials and Methods: Red cells frozen with 40% W/V glycerol and stored at –80°C for up to 37 years were thawed, deglycerolized, and stored at 4°C for 24 h. Results: Red cells frozen for up to 37 years had mean freeze-thaw-wash recovery values of 75%, less than 1% hemolysis, and normal ATP, 2,3-DPG and P50 levels, and 60% of normal RBC K+ levels. Conclusions: Red cells frozen with 40% W/V glycerol can be stored at –80°C for up to 37 years with acceptable in vitro results.

The safety and therapeutic effectiveness of human red cells stored at — 80°C for as long as 21 years

Human red cells frozen by various methods have been stored in the frozen state at —80°C for as long as 21 years. This report discusses: red cells frozen with 42 percent weight per volume (wt/vol) glycerol in an ionic medium in a polyvinylchloride (PVC) plastic bag using the Cohn method; red cells frozen with 45 percent wt/vol glycerol in a low ionic medium in a PVC plastic bag using the Huggins method; red cells frozen with 40 percent wt/vol glycerol in an ionic medium in a polyolefin plastic bag using the Meryman‐Hornblower method; and red cells frozen with 40 percent wt/vol glycerol in an ionic medium in a standard 600‐ml or an elongated 800‐ml PVC plastic primary collection bag with an adapter port using the Naval Blood Research Laboratory (NBRL) method. After frozen storage for as long as 21 years by the four methods described above, the thawed red cells were deglycerolized with 50 to 150 ml of 12 percent sodium chloride and 1.5 to 2.0 l of sodium chloride‐glucose or sodium chloride‐glucose‐phosphate solution. After washing and storage at 4°C for 24 hours, the red cells had a mean freeze‐thaw‐wash recovery value of 90 percent, a mean 24‐hour posttransfusion survival value of 85 percent, a mean index of therapeutic effectiveness of 75 percent, normal or slightly impaired oxygen transport function, and minimal hemolysis. When red cells frozen by the NBRL method in the standard 600‐ml or the elongated 800‐ml primary collection bag for as long as 5.7 years were stored after washing at 4°C for up to 3 days, these units had a mean freeze‐thaw‐wash recovery value of 90 percent, a mean 24‐hour posttransfusion survival value of 85 percent, a mean index of therapeutic effectiveness of 75 percent, normal or slightly impaired oxygen transport function, and minimal hemolysis. Cultures done after storage at 4°C for 1 week showed that the red cells remained sterile. The incidence of container breakage for red cells frozen in the standard 600‐ml or elongated 800‐ml primary collection bag was about 3 percent for units subjected to shipment and less than 1 percent for units that were not transported.

Cryopreservation of Human Platelets Using 6% Dimethyl Sulfoxide and Storage at ‐80°

Platelet studies were done in healthy male volunteers and in thrombocytopenic patients. Some of the platelets used in the study were isolated by mechanical apheresis using either the Haemonetics blood processor 30, the IBM blood processor 2997 or the Fenwal CS‐3000 blood processor before freezing. Other platelets were isolated from individual units of whole blood and pooled before freezing. The platelets were frozen with a 6% cryoprotectant (DMSO) in a polyvinylchloride (PVC) plastic bag or a polyolefin plastic bag at ‐80°C in a mechanical freezer and stored for as long as 3 years. Some of the frozen platelets were transported in dry ice in polystyrene foam containers to determine whether they would be adversely affected by such treatment. Platelet recovery after freezing, thawing and washing was about 75%. In the healthy male volunteers, in vivo recovery of autologous platelets 1–2 h after transfusion was about 33%, and the life span was about 8 days. In the thrombocytopenic patients, in vivo recovery values were 50% of those from fresh platelets. The transfusion of previously frozen washed platelets reduced clinical bleeding in the thrombocytopenic patients with bleeding. There was no evidence of quality deterioration in platelets after storage at ‐80°C for at least 2 years, as determined from in vitro recovery and in vivo survival values, nor was there any adverse effect as a result of shipment of the frozen platelets in dry ice in polystyrene foam containers from one facility to another.

Viability and function of red blood cell concentrates stored at 4 degrees C for 35 days in CPDA-1, CPDA-2, or CPDA-3

The 4OC shelf-life of whole blood and red blood cell concentrates can be extended from 21 days to 35 days by supplementing the CPD anticoagulant with adenine and glucose. In the study reported here, citrate-phosphate-dextrose-adenine (CPDA-1, CPDA-2, and CPDA-3) anticoagulant solutions, were evaluated. CPDA-1 is licensed for red blood cell concentrates with hematocrits of 75 f 3 percent. When CPDA-1 red blood cell concentrates were stored with a hematocrit of 80 f 8 percent for 35 days before autologous transfusion, the mean 24hour posttransfusion survival was an unacceptably low 62 f 11 percent (n =4). CPDA-2 is an experimental anticoagulant, and when used to preserve red blood cell concentrates stored with a hematocrit of 90 f 2 percent for 35 days they showed a mean 24-hour posttransfusion survival value of 74 f 12 percent (n =3). Red blood cell concentrates stored in CPDA-3, also an experimental solution, with an hematocrit of 89 f 1 percent for 35 days, had a mean 24-hour posttransfusion survival of 80 k 8 percent (n = 3). Red blood cell viability was maintained following storage at4OC for 35 days as red blood cell concentrates with hematocrit values of 90 percent in CPDA-2 and CPDA-3, but red blood cell concentrates with hematocrit values of 80 percent stored in CPDA-1 did not maintain viability following 35 days of 4OC storage. After 35 days of 4 C storage, the ATP level of CPDA-1 red blood cell concentrates had decreased to 50 percent of normal, whereas CPDA-2 and CPDAB red blood cell concentrates had 75 percent of normal ATP levels. The 2,3 DPG level, on the other hand, deteriorated faster in CPDA-2 and CPDA-3 red blood cell concentrates during the first 10 days of 4°C storage than in CPDA-1 red blood cell concentrates. After 35 days of 4°C storage, the 2,3 DPG level of CPDA-1, CPDA-2, and CPDA-3 red blood cell concentrates was less than 5 percent of normal. TRANSFUSION 1982;22:210-216.

Autologous platelet-rich plasma isolated using the Haemonetics Cell Saver 5 and Haemonetics MCS+ for the preparation of platelet gel

We compared three methods of isolating platelet‐rich plasma (PRP) using the Haemonetics Cell Saver 5 and one method of isolating PRP by plateletpheresis using the Haemonetics MCS+. PRP contains both platelets and fibrinogen, which are used in the preparation of haemostatic agents.

Correlation between in vitro aggregation and thromboxane A2 production in fresh, liquid-preserved, and cryopreserved human platelets: effect of agonists, pH, and plasma and saline resuspension.

BACKGROUND: Some of the tests used to assess the quality of fresh and preserved platelets (PLTs) include PLT number, PLT morphology, pH of the PLT medium, PLT response to hypotonic stress, and PLT aggregation to agonists. This study was performed to assess the function of fresh and preserved PLTs by their response to aggregation and their production of thromboxane A2 after in vitro stimulation with agonists.

In Vitro Studies of Cryopreserved Baboon Granulocytes

Granulocytes were isolated from the buffy coat of baboon blood by counterflow centrifugation. They were frozen in polypropylene tubes in 2.0 ml volumes containing 1 × 107 granulocytes. The medium consisted of 5% DMSO, 6% HES, 4% human serum albumin, and 6 mM glucose in Normosol‐R, pH 7.1. Granulocytes were cooled to 4 C for 30 minutes, then cooled at 4 C per minute to –80 C and stored for one to three weeks in liquid nitrogen at –197 C. Cooling rates of 1 C and 10 C per minute were less efficacious. Tubes were thawed manually with swirling for 130 seconds at 42 C in a water bath. The yield after thawing was 98 ± 14 per cent.

Circulation and distribution of autotransfused fresh, liquid‐preserved and cryopreserved baboon platelets

Background and Objectives Studies were carried out in five healthy male baboons to determine the 111indium oxine (111In‐oxine) survival of autologous fresh, liquid‐preserved and cryopreserved platelets. Simultaneous organ‐distribution studies were performed to determine the percentage uptake of platelets by the spleen and/or liver.

Survival of baboon biotin-X-N-hydroxysuccinimide and 111In-oxine-labelled autologous fresh and lyophilized reconstituted platelets

Background and Objectivesu2002 In accordance with Food and Drug Administration (FDA) regulations, platelets can be stored in the liquid state at 22 °C for only 5 days. Platelets frozen with 6% dimethylsulphoxide (DMSO) can be stored at −80 °C for 2 years, and platelets frozen with 5% DMSO can be stored at −150 °C for 3 years. Studies are being conducted to determine the effects of lyophilization of platelets. In the present study, we assessed the survival of autologous lyophilized‐reconstituted platelets in the baboon.