The article has summarized partial results of the project of evaluation and implementation method for collection and long-term storage packed red cells (RBC) for strategic, special and therapeutic findings of the Army of the Czech Republic as well as the state. The aim of the study was quality and stability evaluation of cryopreserved RBCs collected by different methods. The source of studied RBCs was double erytrocytapheresis (Group A), whole blood collection with buffy-coat removing (Group B), double erytrocytapheresis with in-line leukofiltration (Group C) and whole blood collection with in-line leukofiltration (Group D). This study compares “non-leukodepleted” RBCs (Group Aand B) and “leukodepleted” RBCs (Group C and D). Tested RBCs were frozen in 40% glycerol and stored at minimum - 65°C for at least 30 days, thawed, deglycerolized, and stored for 21 days at 4±2°C. Glycerolization and deglycerolization were performed with functionally closed system using the Haemonetics ACP 215 machine. In-vitro haematological and biochemical variables were tested on day 0-7-14-21 after deglycerolization. The observed changes of studied haematological and biochemical parameters could be most probably consistent with gradual degradation of non-survival RBCs after freeze-thaw-wash-reconstitution process. Some variables (such as 2,3-DPG, phosphate levels, pH and osmolality) indicate the equilibrium establishment in RBCs during the first 7 days after reconstitution. The results suggest the superior stability of leukodepleted cryopreserved RBCs, collected from whole blood or apheresis, reconstituted in AS-3 after reconstitution related to the non-leukodepleted RBCs, obtained by use of the both methods. It is confirmed by the slow haemolysis and supernatant haemoglobin growth in dependence of storage time. Reconstituted leukodepleted RBCs exhibit significantly higher time stability. No change of 2,3-DPG variable was observed between nonleukodepleted and leukodepleted RBCs. It could be supposed, that this variable is the physiological function of survival RBCs. In contrast to the leukodepletion, the primary source of red blood cells collection (apheresis or whole blood) does not affect stability a quality of RBCs units reconstituted in AS-3. The study demonstrate the superiority of leukodepleted RBCs obtained by apheresis or from whole blood over the non-leukodepleted RBCs (non-filtrated RBCs), reconstituted in AS-3, for extended storage up to 21 days at temperature of 2–6 °C.

        Key words: cryopreservation of red cell, deep frozen blood, reconstitution, glycerolisation, deglycerolisation, crisis blood policy, Nutricel, AS-3, double erythrocytapheresis