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如何在小鼠血小板中进行聚集测定和发光聚集测定。

How to perform aggregometry and lumi-aggregometry in mouse platelets.

机构信息

a Institute for Cardiovascular and Metabolic Research, School of Biological Sciences , University of Reading , Reading , UK.

出版信息

Platelets. 2018 Nov;29(7):638-643. doi: 10.1080/09537104.2018.1478074. Epub 2018 Jun 4.

Abstract

Light transmission aggregometry and lumi-aggregometry are the gold standard platelet assays both clinically and for basic research. The availability of different strains of genetically modified mice and mouse models of human disease means that often laboratories need to use mouse platelets in these assays. Overall, performing aggregometry and lumi-aggregometry with mouse platelets is similar to with human platelets, although methods need be adapted to accommodate their small size, reduced blood volume, and different protein levels. This review aims to highlight these key considerations when planning aggregometry experiments with mouse platelets. These include the method of taking blood, including the use of anticoagulants, as well as the method of platelet preparation, and how to maximize yields. This review also covers how to maximize the number of aggregations that can be performed, both by understanding the minimum requirements of your aggregometer, or by considering new approaches. These include employing high throughput plate-based aggregometry (Optimul), or the use of TPO-mimetics to stimulate platelet production in mice to boost their platelet counts. Finally, phenotypic differences between mouse and human platelets, such as protein expression or sensitivity to agonists are discussed as an important consideration when planning experiments.

摘要

光学比浊法和化学发光比浊法是血小板检测的金标准,无论是在临床还是基础研究中都是如此。由于存在不同遗传修饰品系的小鼠以及人类疾病的小鼠模型,因此实验室通常需要在这些检测中使用小鼠血小板。总体而言,用小鼠血小板进行比浊法和化学发光比浊法与用人血小板进行这些检测非常相似,尽管需要调整方法以适应其较小的体积、减少的血量和不同的蛋白水平。本综述旨在强调在使用小鼠血小板进行比浊法实验时需要考虑的这些关键因素。这些因素包括采血方法,包括抗凝剂的使用,以及血小板的制备方法,以及如何最大限度地提高产量。本综述还涵盖了如何通过了解您的比浊仪的最低要求或考虑新方法来最大限度地增加可进行的聚集次数。这些方法包括采用高通量基于板的比浊法(Optimul),或使用 TPO 模拟物刺激小鼠的血小板生成以提高其血小板计数。最后,讨论了小鼠和人血小板之间的表型差异,例如蛋白表达或对激动剂的敏感性,这是在计划实验时需要考虑的一个重要因素。

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