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

Currie, Laura; Livesey, Steven; Harper, James R.; Connor, Jerome

doi:10.1046/j.1537-2995.1998.38298193098.x

Cited by 39 publications

(47 citation statements)

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“…ThromboSol TM is a platelet‐stabilizing solution consisting of selected second‐messenger effectors that inhibit specific activation pathways endogenous to platelets, resulting in platelets that are biochemically stabilized against the detrimental effects of cold storage (Connor et al , 1996; Currie et al , 1998). Recent studies have demonstrated that the use of ThromboSol in the cryopreservation of platelets allowed simple processing, a reduction of the DMSO to 2% and excellent post‐thaw retention of cell number and in vitro functional activity (Currie et al , 1998). Specifically, when compared to platelets cryopreserved using 6% DMSO, single donor unit platelets cryopreserved with ThromboSol yielded statistically significant higher retention of cell number, percentage of cells displaying a discoid morphology, ESC and HSR (Currie et al , 1998).…”

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confidence: 99%

“…Recent studies have demonstrated that the use of ThromboSol in the cryopreservation of platelets allowed simple processing, a reduction of the DMSO to 2% and excellent post‐thaw retention of cell number and in vitro functional activity (Currie et al , 1998). Specifically, when compared to platelets cryopreserved using 6% DMSO, single donor unit platelets cryopreserved with ThromboSol yielded statistically significant higher retention of cell number, percentage of cells displaying a discoid morphology, ESC and HSR (Currie et al , 1998). In addition, the ThromboSol‐treated platelets displayed a statistically relevant reduction in the expression of the activation marker P‐selectin.…”

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confidence: 99%

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

Currie¹,

Lichtiger

Livesey³

et al. 1999

Br J Haematol

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Summary.Platelet transfusion represents an important component of the therapy for thrombocytopenic patients. Prolonged storage capabilities for platelets would alleviate many problems associated with blood banking. Unfortunately, current cryopreservation methods are complex to implement and result in loss of cell number and functional activity. Previous in vitro studies have shown that the use of ThromboSol TM , a platelet-stabilizing formulation, in the cryopreservation of platelets results in significant retention of cell number and in vitro functional activities in addition to reducing the DMSO requirement to only 2%. We evaluated the in vivo circulatory parameters of platelets cryopreserved with ThromboSol. Single donor platelet units were obtained from healthy volunteers (n ¼ 16); the units were then split and cryopreserved with either ThromboSol and 2% DMSO or 6% DMSO alone. Following storage at ¹80ЊC for 7-10 d the samples were thawed, washed and radiolabelled with either 51 Cr or 111 In. The paired samples were then mixed and reinfused into the autologous volunteer. At various time intervals following transfusion a blood sample was drawn and the quantity of circulating labelled platelets was determined. The percent recovery and survival time was determined by multiple-hit analysis. The ThromboSol-treated platelets, as compared to the 6% DMSO-treated platelets, displayed statistically higher percent recovery (40·2% v 28·8%) and survival time (166·3 h v 152·1 h). These results demonstrated that platelets cryopreserved with ThromboSol displayed superior in vitro and in vivo characteristics as compared to the standard 6% DMSO method. The use of ThromboSol allowed for a 3-fold reduction in the DMSO concentration in conjunction with a 40% increase in circulating cell number and normal survival times.

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confidence: 99%

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

Currie¹,

Lichtiger

Livesey³

et al. 1999

Br J Haematol

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“…Human blood banks typically rely on fresh platelet transfusions prepared from collection of whole blood or apheresis due to the technical challenges of platelet cryopreservation, as well as the considerable loss of platelets and impaired function associated with the freeze/thaw process. 7 As apheresis is not commonly available in veterinary medicine, fresh canine platelets are predominantly collected in whole blood on an as-needed basis. In this study, PC collected via apheresis provided a high-quality product, as evaluated through in vitro functional studies and assessment of in vivo lifespan.…”

Section: Discussionmentioning

confidence: 99%

“…14,19 Expression of P-selectin, a platelet a granule GP exteriorized to the platelet surface following granule release, is considered a marker of platelet activation. 7,20,21 In studies of cryopreserved human platelets, baseline (or unstimulated) P-selectin expression was increased (35%) compared with fresh platelets (9%), suggesting premature platelet activation during the freezethaw-wash process. 17 Cryopreserved human platelets showed reduced P-selectin expression after ADP stimulation, reflecting their decreased ability for normal activation.…”

Section: Discussionmentioning

confidence: 99%

“…In addition, due to the lower in vivo recovery of cryopreserved platelets, it has been estimated that approximately 2.5 U of cryopreserved PCs must be given to achieve a comparable increase in platelet count as for a single unit of fresh PC in thrombocytopenic human patients. 7 Based on the calculated maximum in vivo recoveries of 49.2 and 43.7% for DMSO and Thrombosol PC, respectively (assuming minimal change in recipient TBV following transfusion), the expected increase in recipient platelet count following transfusion of cryopreserved PC ranged from 20,000 to 70,000 platelets/mL. Given the inherent error in automated whole blood platelet count measurements, ie, measured platelet count is generally assumed to be AE20,000/mL, relatively small expected increases in the recipient dogs' platelet counts may not have been detected.…”

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Cryopreservation of Canine Platelets

Appleman

Sachais

Patel

et al. 2009

Veterinary Internal Medicne

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Background: Platelet cryopreservation allows long-term storage and immediate availability of transfusion products. Hypothesis: The addition of a preparation inhibiting platelet activation (Thrombosol, in 2% dimethyl sulfoxide [DMSO]) will enhance in vitro function and prolong in vivo survival of cryopreserved platelets compared with those preserved in 6% DMSO.Animals: Thirty-three research dogs. Methods: Prospective study. Eleven fresh canine apheresis platelet concentrates (PCs) were each split into 3 units: fresh and cryopreserved in 6% DMSO or Thrombosol. Platelet analysis, performed 1-10 weeks postfreezing, included in vitro functional testing and in vivo survival assessed by administration of biotinylated platelets.Results: Platelet aggregation was diminished in cryopreserved PC. Cryopreserved platelets could be activated, as based on mean thrombin-stimulated P-selectin expression (6% DMSO, 23.0%; Thrombosol, 18.4%), although to a lesser extent than fresh PC (49.1%) (P o .0001). The mean maximum in vivo platelet recovery for fresh PC was 80.3%, significantly greater than recovery for 6% DMSO (49.2%) and Thrombosol PC (43.7%) (P .001). The half-life (days) of fresh PC (3.8 AE 0.4) was significantly (P o .002) greater than that of 6% DMSO (1.9 AE 1.0) and Thrombosol (2.4 AE 1.1) PC, with no difference (P 5 .3) between cryopreserved PC.Conclusions and Clinical Importance: Cryopreservation of canine platelets using Thrombosol did not provide any advantage over preservation using 6% DMSO. Cryopreserved platelets can be activated in vitro and provide therapeutic benefit when fresh platelets are unavailable. Further studies are needed to assess their in vivo hemostatic function.

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Genotoxicity related to transfer of oil spill pollutants from mussels to mammals via food

Lemière

Cossu-Leguille

Bispo³

et al. 2004

Environmental Toxicology

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Heavy fuel oils containing high levels of polycyclic aromatic hydrocarbons (PAHs) were released into the marine environment after the Erika oil spill on the Atlantic coast. As highly condensed PAH pollutants can bioaccumulate in invertebrates, their transfer to vertebrates through the food chain was of concern. This study aimed to estimate potential genotoxic effects in rats fed for 2 or 4 weeks with the marine mussel Mytilus edulis contaminated by oil pollutants. Two levels of PAH contamination were studied, around 100 and 500 microg of total PAHs/kg dry weight (d.w.) in mussels. Genotoxic damage in rats was investigated by single-cell gel electrophoresis (Comet assay) and micronucleus assays in liver, bone marrow, and peripheral blood. DNA damage was observed in the liver of rats fed with the most contaminated mussels (500 microg PAHs/kg d.w.).DNA damage also was observed in the bone marrow but less than that in the liver. A small increase in micronuclei frequency was registered as well. This work underlines the bioavailability of pollutants in fuel-oil-contaminated mussels to consumers and the usefulness of the Comet assay as a sensitive tool in biomonitoring to analyze responses to PAH transfer in food. The occurrence of substituted PAHs and related compounds such as benzothiophenes in addition to nonsubstituted PAHs in fuel oils and mussels raised the question of whether they were implicated in the genotoxic effects registered in rats.

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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

Abstract: 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.

Cited by 39 publications

References 36 publications

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

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

Cryopreservation of Canine Platelets

Genotoxicity related to transfer of oil spill pollutants from mussels to mammals via food

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