Research and Development, Australian Red Cross Blood Service, Sydney, New South Wales, Australia.
School of Life Sciences and Proteomics Core Facility, Faculty of Science, University of Technology Sydney, Sydney, New South Wales, Australia.
Transfusion. 2019 Jun;59(6):2093-2102. doi: 10.1111/trf.15204. Epub 2019 Feb 20.
Extending the platelet (PLT) shelf life and enhancing product safety may be achieved by combining cryopreservation and pathogen inactivation (PI). Although studied individually, limited investigations into combining these treatments has been performed. The aim of this study was to investigate the effect of PI treating PLTs before cryopreservation on in vitro PLT quality and function.
ABO-matched buffy coat-derived PLTs in PLT additive solution (SSP+; Macopharma) were pooled and split to form matched pairs (n = 8). One unit remained untreated and the other was treated with the THERAFLEX UV-Platelets System (UVC; Macopharma). For cryopreservation, 5% to 6% dimethyl sulfoxide was added to the PLTs, and they were frozen at -80°C. After being thawed, untreated cryopreserved PLTs (CPPs) and UVC-treated CPPs (UVC-CPPs) were resuspended in plasma. In vitro quality was assessed immediately after thawing and after 24 hours of room temperature storage.
UVC-CPPs had lower in vitro recovery compared to CPPs. By flow cytometry, PLTs demonstrated a similar abundance of GPIX (CD42a), GPIIb (CD41a), and GPIbα (CD42b-HIP1), while the activation of GPIIb/IIIa (PAC-1) was increased in UVC-CPPs compared to CPPs. UVC-CPPs demonstrated greater phosphatidylserine exposure (annexin V) and microparticle shedding but similar P-selectin (CD62P) abundance compared to CPPs. UVC-CPPs displayed similar functionality to CPPs when assessed using aggregometry, thromboelastography, and thrombin generation.
This study demonstrates the feasibility of cryopreserving UVC-PI-treated PLT products. UVC-PI treatment may increase the susceptibility of PLTs to damage caused during cryopreservation, but this is more pronounced during postthaw storage at room temperature.
通过冷冻保存和病原体灭活（PI）相结合，可以延长血小板（PLT）的保存期并提高产品安全性。尽管这两种方法已经分别进行了研究，但对两者联合使用的研究还很有限。本研究旨在研究冷冻保存前用 PI 处理 PLT 对体外 PLT 质量和功能的影响。
从 AB0 匹配的富血小板血浆中提取的 PLTs 在血小板添加剂溶液（SSP+；Macopharma）中混合并分为配对（n=8）。一组不做任何处理，另一组用 THERAFLEX UV-Platelets 系统（UVC；Macopharma）处理。为了冷冻保存，向 PLTs 中加入 5%至 6%的二甲基亚砜，然后在-80°C 下冷冻。解冻后，将未处理的冷冻保存的 PLTs（CPPs）和 UVC 处理的 CPPs（UVC-CPPs）重新悬浮在血浆中。在解冻后立即和室温储存 24 小时后评估体外质量。
UVC-CPPs 的体外回收率低于 CPPs。通过流式细胞术，PLTs 显示出相似数量的 GPIX（CD42a）、GPIIb（CD41a）和 GPIbα（CD42b-HIP1），而 UVC-CPPs 中 GPIIb/IIIa（PAC-1）的激活高于 CPPs。与 CPPs 相比，UVC-CPPs 显示出更高的磷脂酰丝氨酸暴露（annexin V）和微粒释放，但 P-选择素（CD62P）的丰度相似。与 CPPs 相比，UVC-CPPs 在使用聚集仪、血栓弹力图和凝血酶生成评估时表现出相似的功能。
本研究证明了冷冻保存 UVC-PI 处理的 PLT 产品的可行性。UVC-PI 处理可能会增加 PLT 在冷冻保存过程中受损的敏感性，但在室温下解冻后储存时更为明显。
