大容量人血小板冻干保存的初步研究

Preliminary study on the preservation of large volume of human platelets by freeze-drying

目的为使血小板冷冻干燥保存技术用于军队临床战创伤救治,在小容量冻干保存的基础上研究大容量人血小板的冷冻干燥技术。方法采用特制血盒作为冻干容器,以20%(w/v)海藻糖和1%(w/v)牛血清白蛋白作为冻干保护剂。冻干样品复水后测量血小板的数值恢复率和体积分布参数,应用流式细胞仪检测血小板的凋亡率和活化率,采用二磷酸腺苷和肾上腺素检测血小板的聚集功能。结果大容量血小板冻干复水后的数值恢复率为(66.3±2.1)%,平均血小板体积较新鲜血小板有明显增加,冻干血小板的细胞凋亡率达20%左右;冻干和复水过程还造成小部分血小板被激活,其最大聚集率为新鲜血小板的50%左右。结论大容量血小板冻干复水后的恢复率较低,形态结构、活性和功能指标均低于新鲜血小板,但基本可以满足血小板临床止血功能标准。

Objective Towards a clinical application of the technology of freeze-drying preservation of human platelets in certain military situations, a primary study on the freeze-drying of large volume of human platelets was conducted. Methods A specially designed blood vessel was used as the lyophilization container. The human platelets were rehydrated directly after lyophiliztion. The cell recovery rate, mean platelet volume and platelet distribution width were analyzed by a blood-cell counter. The apoptosis rate and activation rate were recorded by a flow cytometry. The aggregation function was tested with adenosine diphosphate and epinephrine. Results The recovery rate of rehydrated platelets reached （66.3±2.1 ） %. Compared with fresh control the mean platelet volume of rehydrated platelets increased significantly. The apoptosis rate was about 20% according to the analysis of the flow eytometry, and a small part of platelets were activated during the freeze-drying and re- hydration process. The maximal aggregation rate was approaching approximately 50% of that of fresh control. Conclusion Freeze-drying preservation of large volume of human platelets resulted in a comparatively low cell recovery rate compared with that of small volumes, and the morphology, aggregation function was also affected, but meet the clinical requirement of coagulation on the whole.