Preparation conditions for optimal Cryoseal quality

Abstract

Introduction: Cryoseal is an autologous fibrin glue automatically prepared using CryoSeal CS-1 system, which generates thrombin and cryoprecipitate (cryo) as two separate components. We investigated how freeze-thawing of plasma, thrombin activity, and fibrinogen concentration affected the quality of Cryoseal components. Methods: Cryoseal was prepared using the CryoSeal CS-1 (Asahi Kasei Medical Corporation). ThromboTrack Solo (Sanko Junyaku Co., Ltd.) was used to measure thrombin activity and clotting time after mixing cryo with thrombin. Fibrinogen concentration and D-dimer were measured on the CN-6000 (Sysmex Corporation). Fresh frozen plasma-LR and pooled plasma were used as source materials. The effects of freeze-thaw cycles on Cryoseal components were assessed by measuring thrombin activity and fibrinogen concentration. The effects of serial dilution of thrombin and cryo on clotting time and clot size were evaluated. Results: Repeated freeze-thawing of plasma minimally affected thrombin activity and fibrinogen concentration. Thrombin activity ≥ 30 U/mL did not affect clotting time after mixing cryo with thrombin. Although there was no apparent change in clotting time with varying fibrinogen concentrations, macroscopic observations showed that higher concentrations of fibrinogen were associated with greater clot firmness and more pronounced clot retraction, possibly due to interactions with other plasma components. Conclusion: To ensure consistent quality of Cryoseal components, thrombin can be prepared in accordance with the manufacturer’s instructions. To obtain cryo with a higher fibrinogen concentration, it is important to carefully remove the excess plasma supernatant from the chamber after cryoprecipitation, thereby concentrating the cryo fraction.