A novel mouse-driven ex vivo flow chamber for the study of leukocyte and platelet function
A novel mouse-driven ex vivo flow chamber for the study of leukocyte and platelet function
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DOI：
10.1152/ajpcell.00500.2003
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发表时间：
2004-04-01
影响因子：
5.5
通讯作者：
Cornelssen, C
中科院分区：
文献类型：
--
作者：
Hafezi-Moghadam, A;Thomas, KL;Cornelssen, C
存在多种用于研究微循环的体外和体内技术。体内系统以其生理保真度令人印象深刻，而体外系统则在可实现的减少量方面表现出色。在这里，我们介绍了自动灌注离体流动室，旨在在明确的血流动力学条件下研究小鼠白细胞和血小板。在我们的模型中，小鼠心脏不断驱动血液流过心室，提供广泛的生理剪切率。我们使用力平衡方法来量化室壁上的主要力。数值模拟显示了基于剪切稀化流体模型的腔室中的流动特性。我们证明了野生型白细胞在固定化 P-选择素上的特异性滚动，这种滚动被阻断性单克隆抗体所消除。当未涂覆时，具有恒定剪切速率的表面支持单个血小板滚动，而在剪切力快速下降的区域，血小板在先前未报道的葡萄状聚集体中相互作用，这表明剪切速率对血小板相互作用类型的影响。总之，离体室相当于连接活体小鼠的动脉和静脉系统的外部容器。该方法结合了白细胞和血小板功能研究中现有体内和体外系统的优点。
Various in vitro and in vivo techniques exist for study of the microcirculation. Whereas in vivo systems impress with their physiological fidelity, in vitro systems excel in the amount of reduction that can be achieved. Here we introduce the autoperfused ex vivo flow chamber designed to study murine leukocytes and platelets under well-defined hemodynamic conditions. In our model, the murine heart continuously drives the blood flow through the chamber, providing a wide range of physiological shear rates. We used a balance of force approach to quantify the prevailing forces at the chamber walls. Numerical simulations show the flow characteristics in the chamber based on a shear-thinning fluid model. We demonstrate specific rolling of wild-type leukocytes on immobilized P-selectin, abolished by a blocking MAb. When uncoated, the surfaces having a constant shear rate supported individual platelet rolling, whereas on areas showing a rapid drop in shear platelets interacted in previously unreported grapelike conglomerates, suggesting an influence of shear rate on the type of platelet interaction. In summary, the ex vivo chamber amounts to an external vessel connecting the arterial and venous systems of a live mouse. This method combines the strengths of existing in vivo and in vitro systems in the study of leukocyte and platelet function.