Viability and Function of Platelets Frozen at 2 to 3C per Minute with 4 or 5 Per Cent DMSO and Stored at −80C for 8 Months

Spector, Jesse I.; Yarmala, J. A.; Marchionni, L.; Emerson, Charles P.; Valeri, C. R.

doi:10.1046/j.1537-2995.1977.17177128894.x

Cited by 37 publications

(27 citation statements)

References 17 publications

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“…In addition to the enhanced recovery of circulating platelets, an important consequence of the use of Thrombo-Sol 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 10year 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.…”

Section: Discussionmentioning

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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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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Section: Discussionmentioning

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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“…Our laboratory has utilized the aspirin‐induced thrombocytopathy to increase the BT and reduce the thromboxane level in the shed blood to assess PLT function of fresh and preserved PLTs in normal volunteers and in baboons 162‐168 . In addition, our laboratory has studied the survival and function of fresh and preserved PLTs in anemic thrombocytopenic patients with prolonged BTs with and without nonsurgical blood loss 169,170 …”

Section: Effects Of Temperature Hct Plt Count and Plt Function On mentioning

confidence: 99%

Nonsurgical bleeding diathesis in anemic thrombocytopenic patients: role of temperature, red blood cells, platelets, and plasma‐clotting proteins

2007

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Research at the Naval Blood Research Laboratory (Boston, MA) for the past four decades has focused on the preservation of red blood cells (RBCs), platelets (PLTs), and plasma-clotting proteins to treat wounded servicemen suffering blood loss. We have studied the survival and function of fresh and preserved RBCs and PLTs and the function of fresh and frozen plasma-clotting proteins. This report summarizes our peer-reviewed publications on the effects of temperature, RBCs, PLTs, and plasma-clotting proteins on the bleeding time (BT) and nonsurgical blood loss. The term nonsurgical blood loss refers to generalized, systemic bleeding that is not corrected by surgical interventions. We observed that the BT correlated with the volume of shed blood collected at the BT site and to the nonsurgical blood loss in anemic thrombocytopenic patients after cardiopulmonary bypass surgery. Many factors influence the BT, including temperature; hematocrit (Hct); PLT count; PLT size; PLT function; and the plasma-clotting proteins factor (F)VIII, von Willebrand factor, and fibrinogen level. Our laboratory has studied temperature, Hct, PLT count, PLT size, and PLT function in studies performed in non-aspirin-treated and aspirin-treated volunteers, in aspirin-treated baboons, and in anemic thrombocytopenic patients. This monograph discusses the role of RBCs and PLTs in the restoration of hemostasis, in the hope that a better understanding of the hemostatic mechanism might improve the treatment of anemic thrombocytopenic patients. Data from our studies have demonstrated that it is important to transfuse anemic thrombocytopenic patients with RBCs that have satisfactory viability and function to achieve a Hct level of 35 vol percent before transfusing viable and functional PLTs. The Biomedical Excellence for Safer Transfusion (BEST) Collaborative recommends that preserved PLTs have an in vivo recovery of 66 percent of that of fresh PLTs and a life span that is at least 50 percent that of fresh PLTs. Their recommendation does not include any indication that preserved PLTs must be able to function to reduce the BT and reduce or prevent nonsurgical blood loss. One of the hemostatic effects of RBC is to scavenge endothelial cell nitric oxide, a vasodilating agent that inhibits PLT function. In addition, endothelin may be released from endothelial cells, a potent vasoconstrictor substance,to reduce blood flow at the BT site. RBCs, like PLTs at the BT site, may provide arachidonic acid and adenosine diphosphate to stimulate the PLTs to make thromboxane, another potent vasoconstrictor substance and a PLT-aggregating substance. At the BT site, the PLTs and RBCs are activated and phosphatidyl serine is exposed on both the PLTs and the RBCs. FVa and FXa, which generate prothrombinase activity to produce thrombin, accumulate on the PLTs and RBCs. A Hct level of 35 vol percent at the BT site minimizes shear stress and reduces nitric oxide produced by endothelial cells. The transfusion trigger for prophylactic PLT transfusion should consider both the H...

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“…12 Cryoplatelets have been transfused in healthy volunteers, generally yielding lower count increments compared to liquid-stored PLTs. 5,13,14 This is caused by biochemical changes during cryopreservation that can be detected in vitro including decreased PLT aggregation, deficient signal transduction, altered morphology, differential expression of certain membrane markers, and microparticle release. [14][15][16][17][18] Recent studies suggest that cryoplatelets are procoagulant, 19,20 but these experiments were performed in static conditions when hemostasis was not restricted by convective hydrodynamic forces.…”

mentioning

confidence: 99%

Comparison between manufacturing sites shows differential adhesion, activation, and GPIbα expression of cryopreserved platelets

Six

Delabie²,

Devreese

et al. 2018

Transfusion

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BACKGROUND Transfusion of cryopreserved platelets (cryoplatelets) is not common but may replace standard liquid‐preserved platelets (PLTs) in specific circumstances. To better understand cryoplatelet function, frozen concentrates from different manufacturing sites were compared. STUDY DESIGN AND METHODS Cryoplatelets from Denver, Colorado (DEN); Sydney, Australia (SYD); and Ghent, Belgium (GHE) were compared (n = 6). A paired noncryopreserved control was included in Ghent. Microfluidic‐flow chambers were used to study PLT adhesion and fibrin deposition in reconstituted blood. Receptor expression was measured by flow cytometry. Coagulation in static conditions was evaluated by rotational thromboelastometry (ROTEM). RESULTS Regardless of the manufacturing site, adhesion of cryoplatelets under shear flow (1000/sec) was significantly (p < 0.05) reduced compared to control. Expression of GPIbα was decreased in a subpopulation of cryoplatelets comprising 45% ± 11% (DEN), 63% ± 9% (GHE), and 94% ± 6% (SYD). That subpopulation displayed increased annexin V binding and decreased integrin activation. PLT adhesion, agglutination, and aggregation were moreover decreased in proportion to that subpopulation. Fibrin deposition under shear flow was normal but initiated faster (546 ± 163 sec GHE) than control PLTs (631 ± 120 sec, p < 0.01), only in the absence of tissue factor. In static conditions, clotting time was faster, but clot firmness decreased compared to control. Coagulation was not different between manufacturing sites. CONCLUSION Cryopreservation results in a subset of PLTs with enhanced GPIbα shedding, increased phosphatidylserine expression, reduced integrin response, and reduced adhesion to collagen in microfluidic models of hemostasis. The proportion of this phenotype is different between manufacturing sites. The clinical effects, if any, will need to be verified.

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Viability and Function of Platelets Frozen at 2 to 3C per Minute with 4 or 5 Per Cent DMSO and Stored at −80C for 8 Months

Cited by 37 publications

References 17 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

Nonsurgical bleeding diathesis in anemic thrombocytopenic patients: role of temperature, red blood cells, platelets, and plasma‐clotting proteins

Comparison between manufacturing sites shows differential adhesion, activation, and GPIbα expression of cryopreserved platelets

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