冰冻干燥(冻干)保存人红细胞在临床输血和战伤救治方面具有重要意义。一些研究表明糖类能提高细胞膜的耐干燥性。试验采用葡萄糖和聚乙烯吡咯烷酮(PVP)作为主要保护剂进行红细胞的冻干保存研究。首先对影响红细胞葡萄糖吸收率的因素,如浓度、孵育温度和时间进行了优化筛选,而后将负载葡萄糖的红细胞与保护液混合并进行冻干保存。结果表明;红细胞对葡萄糖的吸收率依赖于外源葡萄糖浓度、孵育温度以及时间的变化。另外,随着PVP浓度的上升,保护液的结晶起始温度也随之下降,但过度的玻璃化不利于红细胞的冰冻干燥。试验冻干保存的红细胞再水化后的游离血红蛋白浓度低于1g/L。总之,采用PVP和葡萄糖冻干保存红细胞是可行的,但是如何降低PVP对细胞的渗透压伤害是今后研究的重点。
In this study, glucose and PVP were used as main protective reagents to lyophilize human red blood cells. Factors affecting uptake of glucose in red blood cells including extracellular glucose concentration, incubation temperature and time were optimized. Then the mixtures of red blood cells loaded with glucose and protective solutions were lyophilized. The results show that the uptake of glucose was dependent on extracellular glucose concentration, incubation temperature and time. Moreover, the onset temperature of crystal measured by DSC decreased with the increase of PVP concentration, but excessive vitrification may not fit lyophilization of red blood cells. In addition, the free hemoglobin concentration of lyophilize- rehydrated red blood cells was lower than 1g/L. In conclusion, lyophilization of human red blood cells by glucose and PVP is feasible, but how to reduce the osmotic injury of PVP is critical.