人血小板冻干前预处理技术的实验研究

Experimental study on the pretreatment technology of human platelets lyophilization

目的 研究人血小板冻干保存前胞内海藻糖负载技术，筛选出最佳负载海藻糖实验条件。方法 绘制血小板胞内海藻糖负载效率及浓度随温度、时间、胞外海藻糖浓度变化曲线，筛选合适负载条件。结果 海藻糖负载效率与孵育时间（2h后）、温度（30．40℃）呈良好线性关系。在37℃经4h孵育后负载效率可达60％，载入到胞内海藻糖浓度随胞外海藻糖浓度（〈50mM）的升高而递增。结论 37℃、4h为合适负载条件。胞外海藻糖浓度应〈50mM。

Objective To study the technology of loading trehalose into cytoplasm of human platelets before lyophilization, and to determine the optimal experimental conditions of pretreatment technology. Methods The curves of loading efficiency, the internal trehalose concentration versus incubated time, temperature, and external trehalose concentration were drawn up to screen the optimal loading parameters. Results The loading efficiency was linear to the incubated time （2 hours later） and incubated temperature （range from 30℃ to 40℃ ）, respectively. The loading efficiency almost reached 60 percent when the platelets were incubated at 37℃ for 4h. The intracellular trehalose concentration was increased with the raise of the external trehalose concentration （ 〈 50 mM）. Conclusion The optimal incubation temperature and time is 37℃ and 4h respectively, and the concentration of extracellular trehalose should be less than 50 mM.