Metcalfe P, Williamson L M, Reutelingsperger C P, Swann I, Ouwehand W H, Goodall A H
Haematology Division, National Institute for Biological Standards and Control, Potters Bar, Herts.
Br J Haematol. 1997 Jul;98(1):86-95. doi: 10.1046/j.1365-2141.1997.1572983.x.
Three different separation methods, all using centrifugation, are routinely used to prepare therapeutic platelet concentrates from human donor blood. Platelet concentrates derived from platelet-rich plasma (PRP-PC), buffy coat (BC-PC) and apheresis (AP-PC) were investigated at the end of production, and over an 8 d storage period. Change in platelet surface markers were measured by flow cytometry, using fluorescein-conjugated antibodies to fibrinogen, P-selectin (CD62P), GPIIb-IIIa (CD41), GPIb alpha (CD42b) and GPV (CD42d), and fluorescein-conjugated Annexin V was used to measure expression of anionic phospholipid. All concentrates showed some changes during preparation but PRP-PC underwent the greatest changes with significantly higher levels of P-selectin (P<0.001) and bound Annexin V (P=0.001) than AP-PC or BC-PC, and lower levels of GPIb alpha (P=0.002) and GPV (P<0.001). These changes were attributable to component separation rather than venesection. These markers all continued to change on storage with a strong positive correlation between the changes seen during production and those after 5 d storage. PRP-PC continued to show the greatest changes whereas BC-PC showed the least. Fibrinogen was bound to 40-50% of platelets in all preparations and this did not alter significantly on storage whereas total expression of GPIIb-IIIa remained unchanged throughout. There was no evidence that the platelet surface changes were thrombin-mediated and leucocyte depletion of BP-PC by filtration had no effect on the changes. It is proposed that the deterioration of platelet concentrates during storage may be related to activation occurring during preparation. 'Whole blood' flow cytometry using a panel of fluorescein-labelled reagents provides an informative method for evaluating platelet concentrates.
三种不同的分离方法，均采用离心法，常用于从人类献血者血液中制备治疗性血小板浓缩物。在生产结束时以及8天的储存期内，对源自富血小板血浆（PRP-PC）、 Buffy 层（BC-PC）和单采术（AP-PC）的血小板浓缩物进行了研究。使用与纤维蛋白原、P-选择素（CD62P）、糖蛋白IIb-IIIa（CD41）、糖蛋白Ibα（CD42b）和糖蛋白V（CD42d）结合的荧光素抗体，通过流式细胞术测量血小板表面标志物的变化，并使用与膜联蛋白V结合的荧光素测量阴离子磷脂的表达。所有浓缩物在制备过程中均显示出一些变化，但PRP-PC的变化最大，其P-选择素水平（P<0.001）和结合的膜联蛋白V水平（P=0.001）显著高于AP-PC或BC-PC，而糖蛋白Ibα水平（P=0.002）和糖蛋白V水平（P<0.001）较低。这些变化归因于成分分离而非静脉穿刺。这些标志物在储存期间均持续变化，生产期间和储存5天后的变化之间存在强正相关。PRP-PC继续显示出最大变化而BC-PC变化最小。在所有制剂中，40%-50%的血小板结合有纤维蛋白原，并且在储存期间这一比例没有显著改变，而糖蛋白IIb-IIIa的总表达在整个过程中保持不变。没有证据表明血小板表面变化是由凝血酶介导的，并且通过过滤对BP-PC进行白细胞去除对这些变化没有影响。有人提出，血小板浓缩物在储存期间的劣化可能与制备过程中发生的激活有关。使用一组荧光素标记试剂的“全血”流式细胞术为评估血小板浓缩物提供了一种信息丰富的方法。
