冷冻干燥保存兔血小板的实验研究

Experiment on lyophilization of rabbit platelets

目的研究冷冻干燥长期冻干保存的血小板制品的方法。方法将24只大白兔随机分为3个实验组:在所抽取的兔血小板中添加血小板可逆性激活抑制剂、DMSO和海藻糖等低温保护剂,经过预处理、冷冻、一级干燥、二级干燥等过程冷冻干燥兔血小板,分别室温保存1 d(实验1组)、7 d(实验2组)和15 d(实验3组)后经再水化;分别与各自冷冻干燥前的新鲜血小板为对照。并检测血小板回收率和聚集反应功能,并用851Cr示踪法评价血小板体内生存活性。结果3个实验组血小板冷冻干燥后回收率分别为71.68%,68.76%和70.64%,组间无明显差异。实验1、2、3组对凝血酶的最大聚集率(73.20%,77.53%和71.31%)与对照组(86.20%)差异无明显统计学意义(P>0.05),对ADP和没食子酸诱导的聚集反应率(35.6%和58.0%、36.61%和62.56%,32.65%和60.12%)比对照组低(72.4%和91.0%);冻干后保存兔血小板的1、24和48 h体内生存率分别为70%—79%、52%—56%和32%—40%,3组间差异无统计学意义(P>0.05)。结论添加血小板可逆性激活抑制剂、海藻糖和DMSO后冷冻干燥获得的冻干兔血小板,具有较好的聚集活性和体内存活率,保存活性较为稳定。

Objective The aim was to establish a process of platelets lyophilization and find a way of long-term storage of human platelets at room temperature with small size and weight. Methods We divided 24 rabbits into three groups. Reversible platelet activation-inhibitors, DMSO and trehalose were added to rabit platelets. Then the platelets underwent pretreatment, freezing, and first and the second degree desiccation. The platelets were then placed at room temperature. The maximal aggregation, the recovery rate and survival percentage were tested after 1 day （ group 1 ）, 7 days （ group 2 ） and 15 days（ group 3 ） after storage. Results After rehydration, the recovery rates of the three groups were 71.68%, 68.76% and 70. 64%, and there was no difference among them. The thrombin induced maximal aggregations of the rehydrated platelets were not significant different from fresh platelets, but the aggregation induced by ADP or propyl-gallate was decreased. The in vivo Survival rates of the rehydrated platelets were 70%--79% at I hour, 52%--56% at 24 hours and 32%--40% at 48 hours. There was no significantly difference among the three test groups. Conclusion With the addition of reversible activation-inhibitors, DMSO and trehalose, the recovered lyophilized platelets aggregate well and the in vivo survival is satisfactory.