生物化学稳定法冰冻干燥血小板制剂的功能研究

The function of lyophilized platelets stabilized with small molecule carbohydrates

目的对生物化学稳定法——小分子糖稳定法冻干的血小板的功能进行分析研究,进一步验证此种方法保存的血小板的质量。方法采用小分子糖负载的方法对血小板进行预处理保护后冻干;通过细胞计数的方法测定细胞回收率、诱导聚集试验测定血小板冻干制剂的聚集活性、血小板Ⅲ因子有效性试验检测冻干血小板的Ⅲ因子活性;通过动物实验,观察小分子糖稳定法冻干的血小板是否具有缩短实验动物的静脉出血时间的作用。结果小分子糖稳定法冻干的血小板细胞回收率为70%;ADP诱导的聚集活性可达新鲜血小板的50%以上;THR诱导的聚集活性99.9%;缩短实验小鼠的尾静脉出血时间。结论生物化学稳定法冻干的血小板保留了初期止血功能。

Objective To analyze the function of lyophilized platelets stabilized with a biochemical stabilization solution, small molecule carbohydrates stabilization BT40 solution. Methods Small molecule carbohydrate mixture as stabilizer buffer was used for platelet pre-treatment before freeze-drying. The platelet counts of lyophilized and rehydrated platelets were determined by Coulter Cell-DYN 1200. The aggregation activities were tested through SC-2000 Blood Aggregom-eter. Activity of platelet factor 3 were observed. The correction of venous bleeding time of lyophilized platelets was analyzed by Duke method in a mouse model of cyclophosphamide-induced thrombocytopenia.Results Recovery rate of lyophilized platelets protected by small molecule carbohydrate mixture was 〉70%, and most platelets membrane was kept intact. The shape of lyophilized platelets was normal. ADP-induced platelet aggregation was more than 50% of that of fresh platelets. THR induced aggregation was almost with the same as fresh platelets. The activity of platelet factor 3, the structure and function of lyophilized platelets was similar to fresh platelets after 30 days of storage at room temperature.Conclusion The structure and function of membrane glycoprotein GPⅡ/Ⅲand GPⅤ and PF3 are not damaged during platelet lyophilization. Furthermore, animal model experiments showed that lyophilized platelets can shorten the bleeding time after transfusion of rehydrated lyophilized platelets.