降温速率对血小板冰冻保存质量的影响

Influence of cooling rate on quality of cryopreservted platelets

目的:观察不同降温速率下血小板冰冻保存效果,探讨血小板冰冻保存的最佳降温速率。方法:在新型血小板冷冻保护剂作用下,血小板以不同速率(1、10、20℃/min)降温至-80℃后直接转入深低温冰箱保存,冻存1周后复温检测其体外功能和表面分子的变化,与非程序降温组及新鲜血小板作比较。结果:复温后各冰冻保存组的血小板计数均低于新鲜对照组(P<0.05或P<0.01)。肾上腺素诱导的最大聚集率10℃/min组与1℃/min组、非程序降温组两两比较无统计学差异,低于新鲜对照组(P<0.01),但高于20℃/min组(P<0.05)。CD42b表达的平均荧光强度10℃/min组高于其他各冷冻保存组(P<0.05),低于新鲜对照组,但无统计学意义。血小板复温后1℃/min组、20℃/min组、非程序降温组表面的CD62p荧光强度均高于新鲜对照组(P<0.05),10℃/min组CD62p荧光强度高于新鲜对照组,但无统计学差异。结论:降温速率对冰冻保存血小板效果影响较大,在新型血小板冷冻保护剂作用下,相对于1、20℃/min,10℃/min降温速率效果最佳。

Objective：To observe the outcomes of different cooling rates in the cryopreservation of platelets,so as to screen the best cooling rate for cyropreservation of platelets. Methods： Platelet concentrate was cooled down with the new cryopreserving solution at various cooling rates（1,10,20℃/min） and was kept at 80℃ for one week. Then platelets were thawed to check platelet count, adrenaline induced aggregation, and expression of CD41, CD42b and CD62p; the results were compared with those of the non-programmed cooling group and fresh platelets. Results： The platelet counts of eryopreserved groups were lower than that of fresh platelet concentrate（ P〈0.05 or P〈0.01 ）. There were no differences in adrenaline induced aggregation between 10 C/min group, 1 ℃/min group,and non-programmed cooling group; and the aggregation of the former 3 groups was lower than that of 1℃/min group（P〈0. 01） and higher than that of 20℃/min group （P〈0. 05）. The CD42b expression in 10℃/min group was higher than in other groups （P〈0.05） and lower than in fresh platelet concentrate. The CD62p expression in 1℃/min group, 20℃/min group, none programme cooling group was higher than in fresh platelet concentrate（P〈0.05）. The CD62p expression in 10℃/min group was higher than other 3 groups （P〈0.05） and lower than in fresh platelet concentrate, with no statistical significance. Conclusion： Cooling rate has great impact on the quality of cryopreservted platelets. Compared with 1℃/min and 20℃/min, 10℃/min is the optimal cooling rate with the new cryopreservation solution in this study.