Winskel-Wood Ben, Marks Denese C, Johnson Lacey
Research and Development, Australian Red Cross Lifeblood, Sydney, NSW 2015, Australia.
Sydney Medical School, The University of Sydney, Sydney, NSW 2050, Australia.
Int J Mol Sci. 2025 Mar 24;26(7):2944. doi: 10.3390/ijms26072944.
The refrigeration (cold storage) of platelet components provides several benefits over room-temperature (RT) storage, extending the shelf-life up to 21 days. However, the effect of storage conditions on platelet activation in response to stimulation remains unclear. A paired study was conducted where buffy-coat platelet concentrates were pooled, split, and allocated to RT or cold storage (n = 6 in each group). Platelet samples were taken on days 1, 7, 14, and 21, which were tested without stimulation or following activation with TRAP-6, A23187, lipopolysaccharides, or Histone-H4. Imaging flow cytometry was used to assess the surface characteristics of platelets and extracellular vesicles (EVs). The supernatant concentration of EGF, RANTES, PF4, CD62P, IL-27, CD40L, TNF-α, and OX40L was examined using ELISA. Cold-stored platelets generated a greater proportion of procoagulant platelets and EVs than RT-stored platelets in response to stimulation. The supernatant of cold-stored components contained lower concentrations of soluble factors under basal conditions, suggesting that platelet granules were better retained. Cold-stored platelets released higher concentrations of soluble factors following stimulation with TRAP-6, A23187, or Histone-H4. Only cold-stored platelets responded to lipopolysaccharides. These data demonstrate that cold-stored platelets retain the capacity to respond to stimuli after 21 days of storage, which may facilitate improved functional post-transfusion.
与室温储存相比,血小板成分的冷藏(低温储存)有诸多益处,可将保质期延长至21天。然而,储存条件对血小板受刺激后活化的影响仍不明确。开展了一项配对研究,将 Buffy 层血小板浓缩物汇集、分离并分配至室温或冷藏条件下(每组n = 6)。在第1、7、14和21天采集血小板样本,分别在未受刺激或用TRAP-6、A23187、脂多糖或组蛋白-H4激活后进行检测。采用成像流式细胞术评估血小板和细胞外囊泡(EVs)的表面特征。使用酶联免疫吸附测定法检测上清液中表皮生长因子(EGF)、调节激活正常T细胞表达和分泌因子(RANTES)、血小板因子4(PF4)、P-选择素(CD62P)、白细胞介素-27(IL-27)、CD40配体(CD40L)、肿瘤坏死因子-α(TNF-α)和OX40配体(OX40L)的浓度。与室温储存的血小板相比,冷藏的血小板在受到刺激后产生的促凝血血小板和细胞外囊泡比例更高。冷藏成分的上清液在基础条件下可溶性因子浓度较低,表明血小板颗粒保留得更好。在用TRAP-6、A23187或组蛋白-H4刺激后,冷藏的血小板释放的可溶性因子浓度更高。只有冷藏的血小板对脂多糖有反应。这些数据表明,冷藏的血小板在储存21天后仍保留对刺激作出反应的能力,这可能有助于改善输血后的功能。