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用于定量活化绵羊血小板的流式细胞术检测

Flow cytometric assays for quantifying activated ovine platelets.

作者信息

Johnson Carl A, Snyder Trevor A, Woolley Joshua R, Wagner William R

机构信息

Bioengineering Department, University of Pittsburgh, PA, USA.

出版信息

Artif Organs. 2008 Feb;32(2):136-45. doi: 10.1111/j.1525-1594.2007.00498.x. Epub 2007 Nov 14.

Abstract

Ovines are a common animal model for preclinical evaluation of cardiovascular devices including heart valves, endovascular grafts, and ventricular assist devices. Biocompatibility is essential to the success of these devices; however, tools to assess biocompatibility in ovines are limited. To address this need, antibodies that bind to activated human and bovine platelets and annexin V protein were evaluated for potential cross-reactivity to activated ovine platelets. These candidate markers were incubated with stimulated and quiescent ovine whole blood, and binding to platelets was quantified by flow cytometry. Several antihuman CD62P antibodies including one polyclonal antibody, three monoclonal antibodies, and annexin V selectively bound to activated ovine platelets. An assay to quantify platelet microaggregates was also developed. The availability of assays to quantify ovine platelet activation can increase the quality of biocompatibility data obtainable during preclinical development of artificial organs in the ovine model, potentially aiding in the evaluation of design refinements to enhance device biocompatibility.

摘要

绵羊是用于心血管装置临床前评估的常见动物模型,这些装置包括心脏瓣膜、血管内移植物和心室辅助装置。生物相容性对于这些装置的成功至关重要;然而,评估绵羊生物相容性的工具有限。为满足这一需求,对与活化的人及牛血小板和膜联蛋白V蛋白结合的抗体进行了评估,以确定其与活化绵羊血小板的潜在交叉反应性。将这些候选标志物与刺激的和静止的绵羊全血一起孵育,并通过流式细胞术对与血小板的结合进行定量。几种抗人CD62P抗体,包括一种多克隆抗体、三种单克隆抗体和膜联蛋白V,选择性地与活化的绵羊血小板结合。还开发了一种定量血小板微聚集体的检测方法。定量绵羊血小板活化检测方法的可用性可以提高在绵羊模型中人工器官临床前开发期间可获得的生物相容性数据的质量,可能有助于评估设计改进以提高装置生物相容性。

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