Evaluating stored platelet shape change using imaging flow cytometry.
Evaluating stored platelet shape change using imaging flow cytometry.
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使用成像流式细胞术评估储存的血小板形状变化。
DOI：
10.1080/09537104.2022.2136646
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发表时间：
2023
期刊：
影响因子：
3.3
通讯作者：
Stolla,Moritz
中科院分区：
文献类型：
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作者：
Özpolat,Tahsin;Yakovenko,Olga;Stratiievska,Anastasiia;Bailey,SLawrence;Miles,Jeffrey;Usaneerungrueng,Chomkan;Byrne,Daire;Wu,Xiaoping;Stolla,Moritz
血小板在输注前通常在室温下储存5-7天。储存的血小板质量传统上通过Kunicki的形态评分进行评估。这种方法需要大量的培训和经验，并且具有很强的主观性。此外，熟悉这一技术的实验室数量正在减少。血小板的冷藏由于其潜在的优点如减少细菌生长和保存功能，最近重新受到关注。然而，暴露于低温的血小板均匀地从盘状变化为球形，在冷却期间将形态评分结果降低为球形与盘状。我们开发了一种更简单，公正的筛选工具，测量温度诱导的血小板形状变化，使用成像流式细胞术。当减少到两个维度时，球体呈现圆形，而圆盘在从梭形到圆形的光谱上被检测到。我们将圆形事件定义为横轴>纵轴的0.8倍，梭形事件≤纵轴的0.8倍。使用该测定，小鼠和人血小板显示出从梭形到圆形的温度和时间依赖性二维形状变化，与它们从盘到球的三维变化一致。我们在这里描述的方法是一个有价值的工具，用于检测形状变化的差异，响应于激动剂或温度，并将有助于筛选治疗措施，以减轻冷诱导的存储病变。
Platelets are routinely stored at room temperature for 5–7 days before transfusion. Stored platelet quality is traditionally assessed by Kunicki’s morphology score. This method requires extensive training, experience, and is highly subjective. Moreover, the number of laboratories familiar with this technique is decreasing. Cold storage of platelets has recently regained interest because of potential advantages such as reduced bacterial growth and preserved function. However, platelets exposed to cold temperatures change uniformly from a discoid to a spherical shape, reducing the morphology score outcomes to spheroid versus discoid during cooling. We developed a simpler, unbiased screening tool to measure temperature-induced platelet shape change using imaging flow cytometry. When reduced to two dimensions, spheres appear circular, while discs are detected on a spectrum from fusiform to circular. We defined circular events as having a transverse axis of >0.8 of the longitudinal axis and fusiform events ≤0.8 of the longitudinal axis. Using this assay, mouse and human platelets show a temperature and time-dependent, two-dimensional shape change from fusiform to circular, consistent with their three-dimensional change from discs to spheres. The method we describe here is a valuable tool for detecting shape change differences in response to agonists or temperature and will help screening for therapeutic measures to mitigate the cold-induced storage lesion.