Department of Clinical Immunology and Transfusion Medicine (KITM), Karolinska University Hospital, 141 86 Stockholm, Sweden.
Department of Clinical Science, Intervention and Technology (CLINTEC), Karolinska Institutet, 141 52 Huddinge, Sweden.
Int J Mol Sci. 2023 Aug 23;24(17):13097. doi: 10.3390/ijms241713097.
Dimethyl sulfoxide (DMSO) is regularly used as a cryoprotectant agent for the cryopreservation of platelets. However, DMSO is considered toxic. We therefore hypothesized that saline could be used as a non-toxic medium for the cryopreservation of platelets. Double-dose buffy coat platelets ( = 10) were divided and cryopreserved at -80 °C using 5-6% dimethyl sulfoxide (DMSO) or in NaCl (9 mg/mL). Paired testing was conducted pre-freeze, post-thaw (PT 1 h). Upon analysis, each bag was thawed and reconstituted in fresh plasma. Analyses included cell counts and the metabolic, phenotypic, and functional properties of the platelets together with thromboelastometry. The cryopreserved platelets showed several biochemical and ultrastructural changes compared to pre-freezing. Platelet recovery was approximately 17% higher in DMSO-free units ( < 0.001), but the platelet viability was reduced ( < 0.001). However, using controlled freezing ( = 6), the platelet viability was improved. The clot formation time (CFT) was comparable, but DMSO-free platelets showed slightly decreased maximum clot firmness (MCF) ( = 0.034). By reducing the reconstituted plasma volume, a reduced CFT and increased MCF were obtained ( < 0.001). This study demonstrates that platelets can be cryopreserved in saline without the addition of DMSO, with high recovery and maintained hemostatic function. However, controlled freezing is required to optimize platelet quality.
二甲基亚砜（DMSO）常用于血小板的低温保存，作为一种冷冻保护剂。然而，DMSO 被认为是有毒的。因此，我们假设盐水可以作为一种无毒的介质用于血小板的低温保存。双倍剂量的浓缩血小板（= 10）分为两组，分别用 5-6%的二甲基亚砜（DMSO）或氯化钠（9mg/mL）在-80°C下冷冻保存。进行配对测试，包括冷冻前（Pre-freeze）和解冻后 1 小时（PT 1 h）。分析时，每个袋子解冻并在新鲜血浆中重建。分析包括细胞计数以及血小板的代谢、表型和功能特性，同时进行血栓弹性测定。与冷冻前相比，冷冻保存的血小板表现出多种生化和超微结构变化。无 DMSO 单位的血小板回收率约高 17%（<0.001），但血小板活力降低（<0.001）。然而，使用控制冷冻（= 6）可以改善血小板活力。凝血形成时间（CFT）相当，但无 DMSO 血小板的最大凝血硬度（MCF）稍低（= 0.034）。通过减少重建血浆体积，可以获得较短的 CFT 和较高的 MCF（<0.001）。本研究表明，血小板可以在无添加 DMSO 的盐水中冷冻保存，具有较高的回收率和保持止血功能。然而，需要进行控制冷冻以优化血小板质量。
