Improving aspects of platelet cryopreservation would help ease logistical challenges and potentially expand the utility of frozen platelets. Current cryopreservation procedures damage platelets, which may be caused by ice recrystallization. We hypothesized that the addition of a small molecule ice recrystallization inhibitor (IRI) to platelets prior to freezing may reduce cryopreservation-induced damage and/or improve the logistics of freezing and storage. Platelets were frozen using standard conditions of 5-6% dimethyl sulfoxide (Me2SO) or with supplementation of an IRI, N-(2-fluorophenyl)-d-gluconamide (2FA), prior to storage at -80 °C. Alternatively, platelets were frozen with 5-6% Me2SO at -30 °C or with 3% Me2SO at -80 °C with or without 2FA supplementation. Supplementation of platelets with 2FA improved platelet recovery following storage under standard conditions (p = 0.0017) and with 3% Me2SO (p = 0.0461) but not at -30 °C (p = 0.0835). 2FA supplementation was protective for GPVI expression under standard conditions (p = 0.0011) and with 3% Me2SO (p = 0.0042). Markers of platelet activation, such as phosphatidylserine externalization and microparticle release, were increased following storage at -30 °C or with 3% Me2SO, and 2FA showed no protective effect. Platelet function remained similar regardless of 2FA, although functionality was reduced following storage at -30 °C or with 3% Me2SO compared to standard cryopreserved platelets. While the addition of 2FA to platelets provided a small level of protection for some quality parameters, it was unable to prevent alterations to the majority of in vitro parameters. Therefore, it is unlikely that ice recrystallization is the major cause of cryopreservation-induced damage.

中文翻译：

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表征冰重结晶抑制剂改善血小板冷冻保存的能力

改善血小板冷冻保存的方面将有助于缓解后勤挑战，并有可能扩大冷冻血小板的效用。目前的冷冻保存程序会损坏血小板，这可能是由冰重结晶引起的。我们假设在冷冻前向血小板添加小分子冰重结晶抑制剂 (IRI) 可以减少冷冻保存引起的损伤和/或改善冷冻和储存的物流。在 -80 °C 下储存之前，使用 5-6% 二甲亚砜 (Me2SO) 或补充 IRI、N-(2-氟苯基)-d-葡糖酰胺 (2FA) 的标准条件冷冻血小板。或者，在 -30 °C 下用 5-6% Me2SO 或在 -80 °C 下用 3% Me2SO 冷冻血小板，添加或不添加 2FA。在标准条件下 (p = 0.0017) 和 3% Me2SO (p = 0.0461) 但在 -30 °C (p = 0.0835) 下储存后，用 2FA 补充血小板可改善血小板回收率。在标准条件 (p = 0.0011) 和 3% Me2SO (p = 0.0042) 下，2FA 补充剂对 GPVI 表达有保护作用。血小板活化的标志物，如磷脂酰丝氨酸外化和微粒释放，在 -30 °C 或 3% Me2SO 储存后增加，2FA 没有显示出保护作用。尽管与标准冷冻保存的血小板相比，血小板功能在 -30 °C 或 3% Me2SO 下储存后功能会降低，但无论是否使用 2FA，血小板功能都保持相似。虽然向血小板中添加 2FA 为某些质量参数提供了小水平的保护，它无法阻止大多数体外参数的改变。因此，冰重结晶不太可能是冷冻保存引起的损坏的主要原因。