Dumont Larry J, Wolfe Brian, Leite Caroline, Moss Raymond, Wegener Christopher, Thompson Kyle, Min Kyungyoon
Vitalant Research Institute, Denver, Colorado, USA.
Department of Pathology, University of Colorado School of Medicine, Aurora, Colorado, USA.
Transfusion. 2023 Aug;63(8):1554-1562. doi: 10.1111/trf.17464. Epub 2023 Jun 26.
Manufacturing methods for dimethyl sulfoxide (DMSO)-cryopreserved platelets (CPPs) are manual and labor intensive. Thawing and prepare-for-transfusion steps are in an open system that requires transfusion within 4 h. A fill-and-finish system (CUE) can automate the manufacturing process. A newly configured bag system allows freezing, thawing, and use of resuspension solutions while maintaining the functionally closed system, and extending the post-thaw shelf life beyond 4 h. Our objective is to evaluate the feasibility of the CUE system and the functionally closed bag system.
DMSO was volumetrically added to double-dose apheresis platelets, concentrated, and delivered to a 50- or 500-mL ethylene-vinyl acetate (EVA) bag by the CUE (n = 12). The functionally closed bag system contained 25 mL platelet additive solution 3 (PAS-3) in a 50-mL EVA bag. Control CPP (n = 2) were manually prepared. PAS-3 and CPP were thawed together. CPP were stored up to 98 h (20-24°C) and tested using a standard assay panel.
CUE prepared CPP met the design targets: volume, platelet content, and DMSO concentration. CUE CPP P-selectin was high. CD42b, phosphatidylserine (PS) expression, and live cell percentage were favorable compared to controls and favorably maintained over storage. The thrombin generation potency was slightly reduced compared to controls. The 50 mL EVA bag maintained pH for up to 30 h, and the 500 mL EVA bag beyond 76 h.
The CUE system presents a technically feasible method to prepare CPP. A functionally closed bag system with resuspension solution was successful and can extend the post-thaw storage time of CPP.
二甲基亚砜（DMSO）冷冻保存血小板（CPPs）的制造方法是手工操作且劳动强度大。解冻和准备输血步骤是在开放系统中进行，要求在4小时内输血。灌装和成品系统（CUE）可以使制造过程自动化。一种新配置的袋系统允许进行冷冻、解冻以及使用重悬溶液，同时保持功能封闭系统，并将解冻后的保质期延长至4小时以上。我们的目的是评估CUE系统和功能封闭袋系统的可行性。
通过CUE系统（n = 12）将DMSO按体积添加到双倍剂量的单采血小板中，进行浓缩，然后输送到50或500毫升的乙烯 - 醋酸乙烯酯（EVA）袋中。功能封闭袋系统在一个50毫升的EVA袋中含有25毫升血小板添加剂溶液3（PAS - 3）。对照CPP（n = 2）采用手工制备。PAS - 3和CPP一起解冻。CPP在20 - 24°C下储存长达98小时，并使用标准检测面板进行检测。
CUE制备的CPP达到了设计目标：体积、血小板含量和DMSO浓度。CUE CPP的P - 选择素较高。与对照相比，CD42b、磷脂酰丝氨酸（PS）表达和活细胞百分比良好，并且在储存过程中保持良好。与对照相比，凝血酶生成能力略有降低。50毫升EVA袋可将pH维持长达30小时，500毫升EVA袋可维持超过76小时。
CUE系统提供了一种制备CPP的技术可行方法。带有重悬溶液的功能封闭袋系统是成功的，并且可以延长CPP解冻后的储存时间。
