目的探索海藻糖和葡萄糖联合负载红细胞的最适温度,为冷冻干燥红细胞提供基础。方法分别采用0、0.125、0.25、0.5和1mol/L的海藻糖联合葡萄糖在4℃、25℃和37℃负载红细胞6h。然后检测上清液中游离血红蛋白和乳酸脱氢酶含量。结果负载液浓度为1mol/L时,3种温度下的细胞溶血程度较重,即上清中游离血红蛋白和乳酸脱氢酶浓度较高。在浓度低于1mol/L时,25℃负载后,红细胞溶血程度较高。4℃和37℃负载时,上清中游离血红蛋白和乳酸脱氢酶浓度较低。结论在浓度小于1mol/L时,海藻糖联合葡萄糖在4℃和37℃情况下,负载红细胞后对细胞的损伤较小,能够满足冻存的负载要求。
Objective To explore optimal temperature of trehalose and glucose loading red blood cells for lyophilization,red blood cells were incubated with trehalose and glucose at different temperature in the present study.Methods The red blood cells were loaded with 0,0.125,0.25,0.5 and 1 mol/L trehalose and glucose respectively in 4℃,25℃ and 37℃ for 6 hours.Then,the levels of free hemoglobin and lactate dehydrogenase in the supernatants were determined.Results It showed that free hemoglobin and lactate dehydrogenase significantly increased in three groups of 1 mol/L trehalose and glucose(4℃, 25℃ and 37℃).In the trehalose and glucose treatment groups(0.125,0.25,and 0.5 mol/L) at 25℃,the red blood cells showed significant hemolysis.However,red blood cells were loading with 0.125,0.25,and 0.5 mol/L trehalose and glucose at 4℃ and 37℃,the free hemoglobin and lactate dehydrogenase were slightly increased.Conclusion These results suggested that loading with trehalose and glucose(0.125,0.25,and 0.5 mol/L) at 4℃ and 37℃ have little hemolytic effect on red blood cells and be qualified for loading red blood cells for purpose of lyphilization.