Preparation of allogenic RBCs from donor rabbits
Donor rabbits (n =16) were anaesthetised via intravenous injection of 25 mg/kg ketamine and 10 mg/kg xylazine. As described previously, 50 mL/kg of donor blood was withdrawn from the femoral artery17. After removing the platelet-rich plasma and platelet-poor plasma (PPP) from the blood via centrifugation (100 ×g for 15 min), the remaining RBCs were washed with acid citrate dextrose solution. Then, the same volume of mannitol adenine phosphate solution (1.457% [w/v%] D-mannitol, 0.014% adenine and 0.094% sodium dihydrogen phosphate [Terumo, Tokyo, Japan]) was added to prepare allogenic RBCs that were then stored at 4°C. PPP samples showed the following coagulation activity: fibrinogen, 121 mg/dL; AT III activity, 81%; prothrombin time (PT), 13.1 sec; activated partial thromboplastin time (APTT), 31.2 sec, on average.