Hosseini Ehteramolsadat, Beshkar Pezhman, Heydari Farshad, Ghasemzadeh Mehran
Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran.
Department of Laboratory Sciences, School of Allied Medical Sciences, Shahrekord University of Medical Sciences, Tehran, Iran.
Ann Hematol. 2025 May 22. doi: 10.1007/s00277-025-06376-0.
Platelet isolation is a decisive stage in the preparation of platelet concentrate (PC), which may affect functional competence of platelets during storage and post-transfusion. Now considering GPVI as a vulnerable receptor that is mainly affected by shear stress and redox state, this study was conducted for the first time to compare preparation methods of Buffy Coat (BC)- and Platelet-Rich Plasma (PRP)-PCs in terms of GPVI-related phenotypic and functional status. BC- and PRP-PCs were subjected to flow cytometry to analyze GPVI expression and intra-platelet ROS generation. Soluble GPVI were determined by ELISA. Platelet aggregation response to collagen and its adhesion to the collagen matrix were examined by aggregometry and fluorescence microscope, respectively. All the parameters were analyzed on days 0, 1, 3, and 5 of storage. Stored-dependent ROS generation showed significantly higher levels in PRP-PCs compared to BC-PCs on days 1 and 3 of storage. GPVI expression decreased in both products, with BC-PCs showing higher levels on 1 and 3 days of storage. However, with a similar trend, PRP-PCs showed higher levels of shedding, which was significant on day 3. Adhesion/spreading to the collagen matrix also decreased during storage, with higher declines observed in PRP-PCs. Platelet aggregation showed the same pattern with significantly lower PRP-PCs responses to collagen than BC-PCs on the third and fifth days of storage. During platelet storage, GPVI-dependent platelet functional capacity in BC-PCs was better preserved than PRP-PCs, suggesting the priority of BC-PCs method for platelet preparation. In this regard, adopting methods such as platelet isolation and storage in optimal additive solutions, especially those containing ROS scavengers, may help maintain the integrity of GPVI in PRP products. However, further studies, particularly using animal models of thrombus formation, are needed to determine whether the enhanced GPVI function in BC-PCs compared with PRP-PCs can translate into superior efficacy after blood transfusion.
血小板分离是制备血小板浓缩物（PC）的关键阶段，这可能会影响储存和输血后血小板的功能活性。鉴于糖蛋白VI（GPVI）是一个易受剪切应力和氧化还原状态影响的受体，本研究首次比较了富白细胞层（BC）-PC和富血小板血浆（PRP）-PC在GPVI相关表型和功能状态方面的制备方法。对BC-PC和PRP-PC进行流式细胞术分析，以检测GPVI表达和血小板内活性氧（ROS）生成。通过酶联免疫吸附测定（ELISA）测定可溶性GPVI。分别采用凝集测定法和荧光显微镜检查血小板对胶原的聚集反应及其与胶原基质的黏附。在储存的第0、1、3和5天对所有参数进行分析。储存依赖性ROS生成显示，在储存第1天和第3天，PRP-PC中的水平显著高于BC-PC。两种产品中的GPVI表达均下降，BC-PC在储存第1天和第3天显示出较高水平。然而，PRP-PC呈现相似趋势，其脱落水平较高，在第3天具有显著性。在储存过程中，血小板与胶原基质的黏附/铺展也下降，PRP-PC中的下降更为明显。血小板聚集呈现相同模式，在储存第3天和第5天，PRP-PC对胶原的反应显著低于BC-PC。在血小板储存期间，BC-PC中依赖GPVI的血小板功能能力比PRP-PC保存得更好，这表明BC-PC法在血小板制备中具有优先性。在这方面，采用诸如在最佳添加剂溶液中进行血小板分离和储存的方法，尤其是那些含有ROS清除剂的溶液，可能有助于维持PRP产品中GPVI的完整性。然而，需要进一步的研究，特别是使用血栓形成动物模型的研究，以确定与PRP-PC相比，BC-PC中增强的GPVI功能能否转化为输血后更好的疗效。
