冷冻干燥血小板保存技术研究

Lyophilization for Platelet Preservation

为了探索新的血小板保存方法,对血小板进行了冷冻干燥的研究。采用乙醛对血小板做预处理,在冷冻干燥过程中为了稳定血小板的结构,加入了人白蛋白或海藻糖作为保护剂,并对再水化液进行了筛选。对冷冻干燥后的血小板进行了形态学观察和表面膜糖蛋白表达的检测。结果表明,乙醛处理后血小板的回收率基本稳定在60%以上,血小板的聚集活性与处理前相比有轻微的降低;5%人血白蛋白冷冻干燥保护液的保存效果明显优于40mmol/L海藻糖冷冻干燥保护液,在血小板聚集活性保持方面,用贫血小板血浆(PPP)再水化结果明显优于磷酸缓冲盐溶液(PBS)。透射电子显微观察显示,冷冻干燥后血小板胞内细胞器和颗粒物质清晰可见,其中包括线粒体和各种分泌颗粒。乙醛处理后血小板膜糖蛋白的表达与新鲜血小板相比明显降低。结论:乙醛作为一种新的保护剂,对血小板冷冻干燥有良好的效果,值得进一步研究。

To explore a new lyophilized preservation methods for human platelets, platelets were pre-treated with aldehyde, human albumin or trehalose was added to the system of condensed cooling as protectant to stabilize the structure of platelets. The optimal resuspending buffer was also selected in the study. The morphological changes of platelets were observed by using electron microscopy after lyophilization, and the expression of membrane proteins on platelets was detected also after lyophilization. The results indicated that the recovery rate of platelets treated with aldehyde was generally more than 60%. Aggregative ability was reduced a little than the platelet untreated. 5 % of human albumin had an advantage over 40 mmol/L of trehalose in respect of the preservation effect. In the way of keeping aggregative ability, PPP was obviously better than PBS. The results of electron microscopy displayed that organelles including mitochondria and excreted granules could be observed distinctly. Whereas, expression of membrane proteins of platelet treated with aldehyde was evidently dropped as compared with those of the fresh platelet. In conclusion, aldehyde as a novel protective agent, has excellent effects on lyophilization of platelets and is worthy to be further studied.