白膜法汇集浓缩血小板体外质量和功能特性研究

Study on the in vitro quality and function of pooled buffy coat derived platelets

目的研究白膜法汇集浓缩血小板体外质量和功能特性,为国家制定统一的白膜法血小板质量标准提供依据。方法应用白膜法制备汇集浓缩血小板并检测分析各项质量指标,与机采血小板国家标准(GB18469-2001)比对。分别用血小板聚集仪和流式细胞仪测定白膜法汇集浓缩血小板、机采血小板对诱导剂ADP的最大聚集率与血小板膜表面糖蛋白分子CD62P和PAC-1的表达率。结果白膜法制备的汇集浓缩血小板质量指标分别为:容积为(270±14)m l,含量为(2.58±0.51)×1011,pH值为7.24±0.09,红细胞混入量为(6.61±0.16)×109,白细胞混入量为(0.011±0.008)×109。两组血小板对ADP的最大聚集率分别为(85.1±9.0)%和(87.9±11.1)%,P>0.05;两组血小板膜表面糖蛋白分子CD62P表达率分别为(12.3±1.7)%和(11.7±2.1)%,P>0.05;PAC-1表达率分别为(9.3±1.7)%和(8.9±1.4)%,P>0.05。结论白膜法汇集浓缩血小板体外质量指标均达到机采血小板国家标准,血小板功能活性保持良好。

Objective To investigate the in vitro quality and function of pooled buffy coat derived platelets, and to provide the basis for establishment of the national standard for pooled platelets. Methods The platelets were collected by the buffy coat method. The in vitro quality （ platelet yield, residual erythrocytes and leukocytes, volume, and pH value） of the pooled platelets were tested by the KX-21 Sysmex cell counter, the pH monitoring meter, and the ES-1000E thermoanalysis Balance, respectively. Platelet aggregation response to ADP and the activation markers CD62P and PAC- 1 in platelet membranes in the two groups（the pooled platelet group, and the apherosis platelet group ） were detected by APAC3E aggregometry and flow cytometry, respectively. Results The platelet volume, yield, pH value, residual erythrocyte, and residual leukocytes of the pooled platdets were （270 ±14 ） ml, （2.58 ± 0.51 ）×10^11, 7.24±0.09, （ 6.61±0. 16 ）×10^9, and （0.011 ± 0. 008 ）×10^9, respectively. The platelet aggregation response to ADP in the two groups were （85.1 ± 9.0 ） % and （87.9 ±11.1 ） % （ P 〉 0.05 ）. The activation markers CD62P and PAC-1 in platelet membranes of the two groups were （12.3±1.7）% and （11.7±2.1） %（P〉 0.05）, （9.3±1.7）% and （8.9±1.4）% （ P 〉 0.05 ）, respectively. Conclusion The in vitro quality of pooled buffy coat derived platelets can meet the national standard（ GB18469 -2001 ） for apheresis platelet. The function of pooled platelets is normal.