Improved preservation of human red blood cells by lyophilization.
The lyophilization of human red blood cells has important implications for blood transfusion in clinical medicine. In this study, sugars, human serum albumin, polyvinylpyrrolidone, and dimethyl sulfoxide were used as protective reagents for the lyophilization of red blood cells. Freezing temperature, shelf temperature, and the rehydration conditions were optimized. The results showed that extracellular disaccharides, especially trehalose, did not increase the recovery of hemoglobin. However, when the concentration of human serum albumin was higher than 25%, it had a considerable protective effect on the recovery of lyophilized red blood cells; the cellular hemoglobin recovery was over 70%, which was significantly higher than that in the group without human serum albumin (P<0.01). As the concentration of polyvinylpyrrolidone was increased, the extent of vitrification also increased. But when the concentration of polyvinylpyrrolidone was over 40%, the resulting concentration of free hemoglobin was over 1g/L, which was significantly higher than that with 40% (P<0.01). When lyophilization was carried out after freezing at different temperatures, the recovery of cells and hemoglobin was 70-80% and there were no significant differences among the five groups. When the shelf temperature was higher than -30 degrees C, the samples were partly collapsed, but when the shelf temperature was lower than -30 degrees C, the recovery of cells in the -40 and -45 degrees C groups was significantly higher than in the -30 and -35 degrees C groups (P<0.05). The recovery of cells and hemoglobin after lyophilization and rehydration in solutions containing low concentrations of polymers was over 80%, which is significantly higher than the other groups (P<0.01). In addition, when the temperature was higher than 25 degrees C, the concentration of free hemoglobin was significantly lower than it was at 4 degrees C (P<0.01). In conclusion, our study showed the lyophilization of red blood cells is feasible. Disaccharides have no protective effect on lyophilized cells when they are only extracellular and extensive vitrification may be not beneficial. Although the recovery of cells after lyophilization and rehydration by our method was over 70%, the ultrastructure of the cells may be compromised and some hemolysis does still exist. Further research is required.
Han
Ying
Y
Institute of Transfusion Medicine, Beijing 100850, China. hanying1001@yahoo.com.cn
Quan
Guo Bo
GB
Liu
Xiu Zhen
XZ
Ma
En Pu
EP
Liu
An
A
Jin
Peng
P
Cao
Wei
W
D001793:Blood Preservation / Q000379:methods*; D002470:Cell Survival; D015925:Cryopreservation / Q000379:methods; D003451:Cryoprotective Agents / Q000494:pharmacology; D003627:Data Interpretation, Statistical; D004912:Erythrocytes / Q000737:chemistry / Q000166:cytology* / Q000187:drug effects; D005612:Freeze Drying / Q000379:methods*; D006454:Hemoglobins / Q000032:analysis; D006461:Hemolysis / Q000187:drug effects / Q000502:physiology; D006801:Humans; D066298:In Vitro Techniques; D012709:Serum Albumin / Q000494:pharmacology; D013395:Sucrose / Q000494:pharmacology; D014199:Trehalose / Q000494:pharmacology; D014867:Water
D003451:Cryoprotective Agents; D006454:Hemoglobins; D012709:Serum Albumin; D014867:Water; D013395:Sucrose; D014199:Trehalose