Extracellular vesicles in ageing cold-stored whole blood may not compensate for the decreasing haemostatic function in vitro.

Background: Extracellular vesicles (EVs) have procoagulative properties. As EVs are known to accumulate in stored blood products, we compared the EV content and coagulation capacity of leukoreduced cold-stored whole blood (CSWB) with current prehospital and in-hospital component therapies to understand the role of EVs in the haemostatic capacity of ageing CSWB.
Materials and Methods: Blood was obtained from 12 O RhD-positive male donors. CSWB was compared with in-hospital component therapy of red blood cells (RBCs), OctaplasLG and buffy-coat platelets and prehospital component therapy of RBC and lyophilized plasma. Samples were drawn on Days 1 and 14 of CSWB and RBC cold storage. Blood count, haemolysis markers, rotational thromboelastometry, sonorheometry and thrombin generation were analysed. EVs were analysed using nanoparticle tracking analysis and cellular origin was determined using imaging flow cytometry.
Results: There was a trend towards increased production of both platelet and RBC-derived EVs during CSWB storage. Particle count increased during storage, whereas thrombin generation slowed down and in viscoelastic assays, clotting times prolonged, clot formation became impaired, and stiffness of the resulting clot decreased.
Conclusion: Both platelet and RBC-derived EVs increased in number in CSWB during storage. This did not appear to compensate for the in vitro decreasing haemostatic capacity of ageing CSWB, suggesting EVs produced during storage may not have active procoagulative effects, but rather reflect the ageing of blood cells.
(© 2025 Finnish Red Cross Blood Service and The Author(s). Transfusion Medicine published by John Wiley & Sons Ltd on behalf of British Blood Transfusion Society.)