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Abstract
Glycerol and trehalose have been widely examined as protective agents in the cryopreservation of red blood cells (RBCs). However, the effectiveness of these reagents alone on cell viability is moderate. Here, the addition of salidroside attenuated oxidative damage of sheep RBCs prior to and post cryostorage. The supplementation of salidroside to the cryopreservation media containing 10% glycerol improved RBC survival by approximately 61.1±4.8% vs 37.9±4.6%. A smaller effect was seen in RBCs cryopreserved in 300 mM trehalose where the addition of salidroside improved survival by 7.6±0.3%. Furthermore, the addition of salidroside to cold storage solution demonstrated a significant reduction of haemolysis after 4 days for RBCs loaded with either glycerol or trehalose, compared to cells incubated without salidroside. RBCs survival was 2-fold greater following freezing in trehalose, compared with glycerol. After 10 days, salidroside enabled a lower haemolysis of 16.7±1.3% compared to 29.0±8.4% for cells incubated without salidroside. However, salidroside had no effect on RBCs which had been frozen in glycerol as the resulting haemolysis rate by day 10 was approximately 60%. Salidroside increased glutathione reductase activity and decreased lactate dehydrogenase activity. Furthermore, it led to reduced carbonylation of proteins in both glycerol and trehalose loaded cells. Finally, no effect on lipid peroxidation was found in the glycerol loaded RBCs although this was reduced in RBCs loaded with trehalose and salidroside. The present findings confirm the potential use of salidroside as a novel protective agent in cryopreservation and refrigerated storage of sheep RBCs.
Highlights
Red blood cell (RBCs) storage ex-vivo is important for patient care worldwide
The results show that during the incubation conditions, essential preparation step prior freezing, trehalose triggered haemolysis (Fig 1) and has no effect on red blood cells (RBCs) GR and lactate dehydrogenase (LDH) activities (S1C Fig)
Prior freezing sheep RBCs, Sal showed a protective effect in RBCs incubated in trehalose/PP-50 with haemolysis occurring for 2.7±0.6% of the cells, compared to 3.9±1.8% for cells incubated in trehalose/PP-50 without Sal
Summary
Red blood cell (RBCs) storage ex-vivo is important for patient care worldwide. Lost blood must be replaced in major surgeries and RBCs deficiencies must be countered in the treatment of health conditions such as anaemia and sickle-cell diseases. To meet these demands, blood banks are commonplace which use refrigerated storage methods to preserve blood or RBCs. To meet these demands, blood banks are commonplace which use refrigerated storage methods to preserve blood or RBCs This approach only allows blood storage for a maximum of 42 days. The quality of stored RBCs becomes compromised with prolonged storage time [1], rendering these unsuitable for clinical transfusion [2].
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