海藻糖载入红细胞及其冷冻干燥的实验研究

冷冻干燥保存是长期保存人体红细胞的理想方案之一。冻干保护剂海藻糖渗入细胞内后,对细胞膜和细胞内物质有保护作用,其中的一个作用是增加细胞质的浓度,使冻干过程容易形成稳定的玻璃态。应用高渗法处理红细胞,通过考察胞内海藻糖含量、红细胞冻干后的存活率、腺苷三磷酸酶(ATPase)、超氧化物歧化酶(SOD)活力以及细胞形态变化,研究胞内海藻糖含量对红细胞冻干后活性的影响。结果显示:海藻糖对红细胞冻干具有明显的保护作用,随胞内海藻糖浓度升高,其保护性能逐渐增强;43.8mmol/L的胞内海藻糖浓度对红细胞保护最好,细胞存活率达到53.6%,形态保持良好,ATP和SOD活力均在正常的范围内。

Effects of Intracellular Trehalose on the Freeze-Dried Red Blood Cells

Freeze-drying is one potentially ideal technology for long-term preservation of living biological cells. To increase the survival and stability of human red blood cells (RBCs) after freeze-drying and rehydration, trehalose is introduced in RBCs using a hypertonic method before freeze-drying. The effects of intracellular treha- lose concentration on RBCs after freeze-drying and rehydration are investigated. The results indicate that the sur- vival of RBCs after freeze-drying and rehydration increases with the increment of intracellular trehalose concentration, the survival of RBCs after freeze-drying and rehydration is over 53.6% with 43.8 mmol/L of intracellular trehalose, the levels of ATPase and superoxide dismutase (SOD) are maintained close to the levels of fresh RBCs. Morphologi- cal study also confirms the results. It demonstrates that disiccation tolerance of RBCs increases along with the concentration of intracellular trehalose. Our study demonstrates that trehalose has protective effects on freeze-dried RBCs, the survival of freeze-dried RBCs depends on both extracellular and intracellular vitrification.