目的:探索海藻糖和葡萄糖联合负载红细胞的最适时间,为冷冻干燥红细胞提供基础。方法:分别采用0、0.125、0.250、0.500和1.000mol/L的海藻糖联合葡萄糖在37℃负载红细胞4、6和8h。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种负载时间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,负载时间为4、6h时,上清中游离血红蛋白和乳酸脱氢酶浓度较低。而负载时间延长至8h时,红细胞溶血程度较高。结论:在浓度小于1mol/L时,海藻糖联合葡萄糖在37℃情况下负载红细胞6h时对细胞的损伤较小,能够满足冻存要求。
Objective:To explore the optimum time of trehalose and glucose loading red blood cells for tyophilisation. Method:The red blood cells were loaded with 0,0. 125,0. 250,0. 500 and 1. 000 mol/L trehalose and glu- cose respectively in 37℃ for 4,6, and 8 hours. Then, the levels of free hemoglobin and lactate dehydrogenase in the supernatants were determined. Result: The results showed that free hemoglobin and lactate dehydrogenase signifi- cantly increased in three groups of 1mol/Ltrehalose and glucose(4,6, and 8 h). When red blood cells were loading with 0. 125,0. 250, and 0. 500 mol/L trehalose and glucose for 4 and 6 h, the free hemoglobin and lactate dehydrogenase were low. However, when the incubation time extend to 8h(0. 125,0. 250,and 0. 500 mol/L),the red blood cells showed significant hemolysis. Conclusion : These results suggested that loading with trehalose and glucose (0. 125,0. 250 ,and o. 500 mot/L)at 37℃ for 6h had little hemolytic effect on red blood cells and were qualified for loadiflg red blood cells for purpose of lyphilization.