冰冻血小板透析式洗涤方法研究

Protocol of dialysis-type processing on frozen platelets

目的建立透析式洗涤冰冻血小板的技术方法,并检测血小板的功能指标,对透析式洗涤效果进行评价。方法样品为机采血小板1 U/袋,共20袋,分为离心处理组和透析处理组,通过血小板体外功能检测方法,观察冰冻血小板的洗涤效果。结果血小板经两种方法处理后,透析组洗涤后的回收率[(73.57±12.49)%]明显优于离心组[(62.21±7.07)%],其渗透压值[(305.8±5.27)m Osm/kg]低于离心组[(383.2±22.00)m Osm/kg],透析组几乎能将二甲基亚砜(DMSO)全部去除,而离心组DMSO残留量约为0.8%,两组间差异有统计学意义(P<0.05)。透析组和离心组平均血小板体积[(9.44±0.93)vs(9.23±0.50)fl]、ADP诱导聚集率[(20.7±8.00)%vs(27.7±11.83)%]、凝血酶诱导血小板聚集率[(84.1±7.88)%vs(92.9±5.28)%]、低渗休克反应[(19.02±4.42)%vs(23.19±9.08)%]、活化率[(16.00±9.40)%vs(21.86±8.14)%]和凋亡率[(46.31±10.28)%vs(54.59±16.76)%],经两法处理后差异无统计学意义(P>0.05)。结论与离心处理洗涤方法相比,该课题组自主研发的冰冻血小板透析式洗涤机不仅能提高血小板回收率,也有效解决了DMSO残留量过多的问题。

Objective To establish a new dialysis washing method for human frozen platelets and to evaluate a new type of washing machine based on dialysis.Methods Twenty bags of platelets（1 U） were divided into two groups：centrifugation group and dialysis group.The effect of the washing machine was evaluated by measuring platelet function in vitro.Results After the platelets were treated by the two methods,the recovery rate of dialysis group [（73.57 ±12.49） %] was significantly better than that of centrifugal group [（62.21 ± 7.07） %].The osmotic pressure of dialysis group was 305.8 ± 5.27 m Osm/kg,compared to 383.2 ± 22.00 m Osm/kg in centrifugation group.There was significant differences in the recovery rate and osmotic pressure（P〈0.05）,but there was no significant difference between dialysis group and centrifugation group in the mean platelet volume [（9.44 ± 0.93） vs（9.23 ± 0.50） fl ],ADP-induced aggregation rate [（20.7 ± 8.00） % vs（27.7 ± 11.83） % ],thrombin-induced platelet aggregation rate [（84.1 ± 7.88） %vs（92.9 ± 5.28） % ],hypotonic shock responses [（19.02 ± 4.42） % vs（23.19 ± 9.08） % ],CD62 p activation rate[（16.00 ± 9.40） % vs（21.86 ± 8.14） %] or apoptosis rate [（46.31 ± 10.28） % vs（54.59 ± 16.76） %]（P〈0.05）.Conclusion Compared to centrifugation group,the frozen platelet washing machine can not only improve platelet recovery,but also solve the problem of excessive DMSO residues.