Mangin Pierre H, Gardiner Elizabeth E, Nesbitt Warwick S, Kerrigan Steven W, Korin Netanel, Lam Wilbur A, Panteleev Mikhail A
INSERM, EFS Grand-Est, BPPS UMR-S1255, FMTS, Université de Strasbourg, Strasbourg, France.
ACRF Department of Cancer Biology and Therapeutics, The John Curtin School of Medical Research, The Australian National University, Canberra, ACT, Australia.
J Thromb Haemost. 2020 Mar;18(3):748-752. doi: 10.1111/jth.14717.
Experimental videomicroscopic in vitro assays of thrombus formation based on blood perfusion are instrumental in a wide range of basic studies in thrombosis, screening for hereditary or acquired plateletrelated pathologies, and assessing the effectiveness of novel anti-platelet therapies. Here, we discuss application of the broadly used "in vitro thrombosis model": a frequently used assay to study the formation of 3D aggregates under flow, which involves perfusing anticoagulated whole blood over fibrillar collagen in a flow geometry of rectangular cross-section, such as glass microcapillaries or parallel-plate flow chambers. Major advantaged of this assay are simplicity and ability to reproduce the four main stages of platelet thrombus formation, i.e. platelet tethering, adhesion, activation and aggregation under a wide range of hemodynamic conditions. On the other hand, these devices represent, at best, simple reductive models of thrombosis. We also describe how blood flow assays can be used to study various aspects of platelet function on adhesive proteins and discuss the relevance of such flow models. Finally, we propose recommendations for standardization related to the use of this assay that cover collagen source, coating methods, micropatterning, sample composition, anticoagulation, choice of flow device, hemodynamic conditions, quantification challenges, variability, pre-analytical conditions and other issues.
基于血流灌注的血栓形成实验性视频显微镜体外检测方法，在血栓形成的广泛基础研究、遗传性或获得性血小板相关疾病筛查以及评估新型抗血小板疗法的有效性方面发挥着重要作用。在此，我们讨论广泛使用的“体外血栓形成模型”的应用：这是一种常用的检测方法，用于研究在流动条件下三维聚集体的形成，该方法包括在矩形横截面的流动几何结构（如玻璃微毛细管或平行板流动腔）中，将抗凝全血灌注在纤维状胶原蛋白上。该检测方法的主要优点是简单，并且能够在广泛的血流动力学条件下重现血小板血栓形成的四个主要阶段，即血小板 tethering、粘附、活化和聚集。另一方面，但这些装置充其量只是血栓形成的简单简化模型。我们还描述了血流检测如何用于研究血小板在粘附蛋白上功能的各个方面，并讨论了此类流动模型的相关性。最后，我们针对该检测方法的使用提出标准化建议，涵盖胶原来源、包被方法、微图案化、样品组成、抗凝、流动装置的选择、血流动力学条件、定量挑战、变异性、分析前条件及其他问题。
