电渗透用于强化海藻糖载入红细胞的实验研究

Experiment study of loading trehalose into human red blood cells by electropermeabilization

海藻糖是一种有效的冻干保护剂,实验采用电渗透的方法强化海藻糖载入红细胞,在不同电压、脉宽、频率和细胞外海藻糖浓度的条件下对红细胞进行电渗透.实验结果表明,当电压300 V、脉宽1 ms、频率4次/min、细胞外海藻糖浓度800 mmol/L时,细胞内海藻糖浓度达到(63.68±2.14)mmol/L.电渗透后红细胞形态正常,细胞膜的渗透脆性略有降低,电渗透致溶血率为17%,ATP酶活性与新鲜红细胞无显著差异.

Electropermeabilization was adopted to load trehalose into red blood ceils in the investigation. Human red blood cells were electropermeabilized under different conditions of electric voltage, pulse width, frequency and extracellular trehalose concentration. The results show that intracellular trehalose concentration reaches （63.68 ± 2.14）mmol/L under the condition of 300 V, 1 ms, 4 pulses/min and 800 mmol/L extracellular trehalose concentration. Mter electropermeabilization,the red blood cells retain normal morphology, the membrane osmotic fragility decreases slightly, the hemolysis rate due to electropermeabilization is 17% and the ATP enzyme activity is almost similar to that of fresh red blood cells.