Objective To investigate whether the biochemical function and morphology of stored outdated erythrocytes were able to be restored to normal levels.Methods The experiments were carried out with 4 groups of erythrocytes. Group 1 consists of post-rejuvenated erythrocytes, which were harvested by incubating twice washed stored erythrocytes on the 3 rd day after outdate at 37℃ for 1h, with a rejuvenating solution containing adenine 5mmol/L, inosine 100mmol/L, pyruvate 100mmol/L, phosphate 103mmol/L. Group 2 was pre-rejuvenate erythrocytes, obtained by washing stored erythrocytes on the 3rd day after outdate. Group 3 was saline control erythrocytes, which were incubated with 0.9% saline. Group 4 was freshly collected erythrocytes used as normal controls. ATP, 2, 3-DPG, morphology scores, MCV, osmotic fragility, and hemoglobin content of RBC in four groups were measured.Results The post-rejuvenate RBC ATP [(3.41?0.52)?mol/gHb] was significantly higher(P0.05) compared with fresh RBC [(3.45?0.51)?mol/gHb].The post-rejuvenate RBC 2,3-DPG [(14.08?3.10)?mol/gHb] was significantly higher(P0.05) compared with fresh RBC [(14.87?3.10)?mol/gHb].The post-rejuvenation morphology score(130.00?10.80) was significantly greater (P0.05) compared with fresh RBC (120.00?5.00). The post-rejuvenate mean corpuscular volume [(89.30? 7.20)fl] was significantly lower (P0.05). The hemoglobin shed in vesicles during rejuvenation was significantly greater than that in the saline control [(0.51?0.33)vs(0.20?0.18)mg/dl RBCs;P