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

Abstract

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.

Enhanced circulatory parameters of human platelets cryopreserved with second‐messenger effectors: an in vivo study of 16 volunteer platelet donors

“…In addition to the enhanced recovery of circulating platelets, an important consequence of the use of ThromboSol is the ability to reduce the requirement for DMSO as a cryoprotectant. Previous studies testing the use of reduced concentrations of DMSO during infusion indicated that the recipients showed no adverse effects, even following a 10‐year period (Handin & Valeri, 1972; Lazarus et al , 1981; Melaragno et al , 1985; Spector et al , 1977). The development of a cryopreservation system that contains transfusable components would eliminate both the damaging effects of the wash step and the need to break the closed system.…”

“…This loss of in vitro functional activity is also reflected in the cryopreserved platelets' in vivo circulatory parameters (Daly et al , 1979; Handin & Valeri, 1972; Kim & Balduini, 1974; Melaragno et al , 1985; Schiffer et al , 1976; Spector et al , 1977). Following infusion, the recovery of 51 Cr‐labelled cryopreserved platelets ranges from 30% to 40% as assessed in multiple studies (Handin & Valeri, 1972; Melaragno et al , 1985; Valeri, 1974).…”

“…This loss of in vitro functional activity is also reflected in the cryopreserved platelets' in vivo circulatory parameters (Daly et al , 1979; Handin & Valeri, 1972; Kim & Balduini, 1974; Melaragno et al , 1985; Schiffer et al , 1976; Spector et al , 1977). Following infusion, the recovery of 51 Cr‐labelled cryopreserved platelets ranges from 30% to 40% as assessed in multiple studies (Handin & Valeri, 1972; Melaragno et al , 1985; Valeri, 1974). Taken in conjunction with the loss of cell numbers as a consequence of the freeze–thaw process and the post‐thaw wash step, the final in vivo recovery can be as low as 18% of the original fresh platelet population (Spector et al , 1977; Valeri et al , 1974a, b).…”

Reduction of blood loss by infusion of human platelets in a rabbit kidney injury model

“…Researchers are experimenting by treating platelets with dimethyl sulfoxide and freezing them in liquid nitrogen 9,10 . Cryo‐preservation of platelets extends the shelf life of platelets to 1 year but requires extensive washing and processing to remove the cryoprotective agents 11 . Another approach to platelet preservation has been to lyophilize the platelets.…”

Novel treatment modalities: new platelet preparations and subsititutes

“…Recovery post‐thaw is approximately 75% and after transfusion into normal volunteers, in vivo platelet recovery at 1 h is approximately 33% with a platelet survival thereafter of about 8 d (Melaragno et al , 1985). Cryopreserved platelets acquire a variety of morphological and functional defects in vitro (Spector et al , 1977; Shepherd et al , 1984; Melaragno et al , 1985) but are haemostatically effective in vivo (Schiffer et al , 1978; Melaragno et al , 1985). In one recent study, cryopreserved platelets were reported to be more effective in decreasing blood loss in cardiopulmonary bypass surgery than standard liquid stored platelets (Khuri et al , 1999).…”