Determining the effects of varying blood storage conditions on clot formation and digestion under shear

Studies aiming to understand the effects of storage on whole blood clotting often rely on characterizing coagulation under static conditions. Minimal work has explored the effects of physiologic shear on clot formation and thrombolysis utilizing fractionated and reconstituted whole blood products. Whole blood (WB) was fractionated into platelet free plasma (PFP), packed red blood cells (pRBCs), and platelets storing each component under its ideal conditions - including platelet cryopreservation. Recombination at their native ratios was accomplished over 91-days of storage and clotting/thrombolysis was analyzed utilizing thromboelastography (TEG) and Chandler loop. Reconstituted whole blood (rWB) preserved clot strength through 91 days with minimal deviation from baseline, in contrast to WB stored at 4°C which experienced a significant decline by storage day-42. Clot formation under shear for both rWB and WB led to increased clot mass through storage. No significant deviation from baseline was appreciated until day-70 of storage in rWB. Increasing degrees of thrombolysis were seen in both groups, with rWB significantly deviating from baseline at day-70. No significant changes in overall clot architecture occurred throughout storage and recombination. This fractionation and recombination protocol serves as a method to further develop reproducible in vitro clot analogs for preclinical thrombolytic therapy screening.