【作者】 孟凡会； 孙青刚； 孙波； 刘竹珍； 曹永献； 赵洪国；

【Author】 Meng Fanhui, Sun Qinggang, Sun bo, et al Medical School Hospital of Qingdao University, 266003

【机构】 青岛大学医学院附属医院；

【摘要】 目的观察体积分数0．02二甲亚砜(DMSO)联合第二信使效应剂(ThromboSol)在-196 ℃时对浓缩血小板膜CD42b、CD62p表达的影响,并了解体积分数0．02DMSO+ThromboSol(TS) 对浓缩血小板的长期(6个月)保存效果。方法分别应用体积分数0．02DMSO、体积分数 0．06DMSO、体积分数0．02DMSO+ThromboSol做为冻存液在-196℃冷冻保存浓缩血小板,并在冻存后1、3及6个月时分别取出复温,流式细胞仪检查血小板膜CD42b、CD62p表达水平。结果冻存后1、3及6个月,体积分数0．02DMSO+TS组与体积分数0．06DMSO组血小板CD42b 的表达水平差异无显著性(q=1．12-1．76,P>0．05),两者均高于体积分数0．02DMSO组(q=3．17-4．06, P<0．0),但与新鲜组相比,3组冷冻保存血小板CD42b表达水平均有所下降(F=3．13-5．01, q=3．09-4．15,P<0．05)。体积分数0．02DMSO+TS组CD62p的表达水平低于体积分数0．06DMSO 组和体积分数0．02DMSO组,差异有显著性(q=3．20-4．34,P<0．05),后两组差异无统计学意义 (q=1．55-2．02,P>0．05),与新鲜组相比,3组均明显升高,差异有显著性(F=5．75-9．77,q=4．85-7．35, P<0．01)。结论第二信使效应剂的应用降低了DMSO的使用浓度,在抑制血小板的体外激活方面,体积分数0．02DMSO+ThromboSol的效果优于体积分数0．06DMSO,且长期(6个月)保存效果稳定。更多还原

【Abstract】 Objective To observe the effect on platelet membrane CD42b and CD62p after cryopreservation using 0.02% dimethyl sulfoxide(DMSO) plus ThromboSol, a mixture of second -messenger effectors, and to realize the long - term (6 months) protective function of this platelet storage solution.Methods Platlet concentrates were treated with 0.02% DMSO,0.06%DMSO and 0.02%DMSO plus ThromboSol, then placed in liquid nitrogen.After frozen for 1 month, 3 months or 6 months, samples were thawed and expression levels of CD42b and CD62p on platelet membrane analyzed.Results According to the results of platelet expression levels of CD42b, platelets treated with 0.02DMSO plus ThromboSol were similar to those stored by the method using 0.06% DMSO after cryopreseved for lmonth,3 months and 6months(q=1.12-1.76, P>0.05).They were all superior to those frozen with 0.02% DMSO(q=3.17-4.06, P<0.05).In comparison to the fresh platelets, three groups of frozen platelets showed significantly decrease (F=3.13-5.21, q=3.09-4.15, P<0.05).The platelets treated with 0.02%DMSO plus ThromboSol showed statistically lower CD62p expression than those frozen with 0.06%DMSO and 0.02% DMSO (q=3.20-4.34,P<0.05).There were no obviously CD62p expression differences between platelets treated with 0.06%DMSO and 0.02%DMSO (q= 1.55-2.02, P>0.05).But three groups of frozen platelets showed significantly increase of surface expression of CD62p in comparison to fresh platelets (F=5.75-9.77, q=4.85-7.35, P<0.01).Conclusion The use of second-messenger effectors could reduce the DMSO concentration from 6 to 2 percent.The CD62p membrane expressive levels showed that the group of 0.02%DMSO plus ThromboSol is suprior to the group 0.06%DMSO in inhibiting the activation of pletelets.And this cryopreservative method was stable within 6 months.更多还原