Synergistic Effect of Trehalose and Saccharose Pretreatment on Maintenance of Lyophilized Human Red Blood Cell Quality

Yan-Qiong Li,Li-Hui Zhong,Rui Hu,Qian Sun,You-Ping Yan

doi:10.4314/tjpr.v15i3.14

Abstract

Purpose: To investigate the synergistic effect of trehalose and saccharose pretreatment on maintenance of lyophilized human red blood cell (RBC) quality. Methods: RBCs were pre-treated with trehalose and saccharose, and then lyophilized and re-hydrated. Prior to lyophilization and after re-hydration, RBC parameters, RBC counts, total hemoglobin concentration, mean corpuscular hemoglobin (MCH), mean corpuscular volume (MCV), comprehensive deformation index, hemolysis ratio and phosphatidylserine (PS) expression, were determined using a hematology analyzer, an RBC deformation instrument, a spectrophotometer and a flow cytometer, respectively. Superoxide dismutase (SOD), glucose-6-phosphate dehydrogenase (G-6-PD), and adenosine triphosphatase (ATPase) activities were determined using kits for SOD, ATPase, and G-6PD assay, respectively. Results: After lyophilization-rehydration, RBC counts and total hemoglobin recovery rates, deformability, and RBC SOD, ATPase, and G-6-PD activities were significantly decreased by 47.24 – 74.65 % (p 0.05). Trehalose and saccharose pretreatment significantly reversed the effects of lyophilization-rehydration on these RBC parameters by approximately 13.16 – 211.11 % (p < 0.01), compared with the control group. The combined effects were synergistic. Conclusion: Trehalose and saccharose pretreatment synergistically enhances maintenance of lyophilized RBC quality.

Highlights

Human red blood cell (RBC) preservation methods include short-term preservation at 4 °C for 35 – 42 days and long-term preservation at 80 or -196 °C for approximately 10 years
Effects of trehalose and saccharose pretreatment on conventional lyophilized RBC indicators After lyophilization-rehydration, the RBC counts and total hemoglobin recovery rates in the TG, SG, and TSG were significantly increased by 21.24 - 180.57 % (p < 0.01), compared with the
The lyophilized RBC comprehensive deformation index in the CG was significantly decreased by 56.25 % (p < 0.01), compared with the NG (Fig. 2)

Summary

INTRODUCTION

Human red blood cell (RBC) preservation methods include short-term preservation at 4 °C for 35 – 42 days and long-term preservation at 80 or -196 °C for approximately 10 years. Compared with the short- and long-term methods, lyophilization advantages include longterm room temperature preservation, convenient transportation, and simple storage conditions [2,3]. Despite these advantages, problems such as membrane damage, hemoglobin leakage, and high RBC hemolysis ratio have been reported [4]. The 20 mL 45 % hematocrit RBCs samples were separately pre-treated with different concentrations of trehalose and saccharose as described above. Each sample of pre-treated RBCs (0.3 mL) was re-suspended in 0.7 mL lyophilization buffer and transferred into a freeze dry vial and equilibrated for 30 min at 4 °C. Hb2 and Hb1 were total hemoglobin concentrations before lyophilization and after rehydration, respectively

Evaluation of RBC deformability

RESULTS

DISCUSSION

CONCLUSION

CONFLICT OF INTEREST