体外血小板流动腔研究方法的建立和应用

The Development and Application of A Flow Chamber for Platelet Research

目的建立模拟体内血液流动环境下,实时观察、研究血小板与血管表面血管性假性血友病因子(vWF)相互作用的模型。方法利用包被vWF的细玻璃管模拟血管,分别采用负压装置、蠕动泵和恒流注射泵产生不同流态和精确剪切率的流体环境,结合倒置显微镜和电荷耦合器件图像传感器(CCD,Charge-coupleddevice)实时观察和记录血小板或细胞在流体作用下的一系列反应。结果血小板和转染了血小板糖蛋白Ib-Ⅸ的中国仓鼠卵巢细胞(Chinese hamster ovary,CHO),在不同流态下都可与玻璃管壁上的vWF相互作用,从而介导血小板或CHO细胞与玻璃管壁的瞬时结合(滚动),并最终牢固结合在玻璃管壁上。结论本研究方法可以在精确剪切率和不同流态的流体环境条件下,实时地观察、记录血小板与vWF相互作用的情况,为血小板各种生理功能的研究以及相关药物的研究应用创造条件。

Objective To establish an in vitro model for observing the real-time interaction between platelets and vascular vessel. Methods Thin glass tubes coated with von Willebrand faeter （vWF） were used to form the flow chambers. Siphonage system, peristaltic pump and a constant flow syringe pump were adopted to supply different kinds of flow and well defined shear rates. With inverted microscope and charge- coupled device （CCD） ,a series of reactions of platelets and chinese hamster ovary （CHO） cells transfeeted with glycoprotein Ⅰb-Ⅸ under flow conditions were recorded simultaneously. Results Platelets and CHO cells transfected with glycoprotein Ⅰb-Ⅸ rolled and steadily adhered on the tube wall through binding to vWF under different kinds of flow and well defined shear rates. Conclusion This flow chamber allows real- time observation and record of the interaction between platelets and vWF under well-defined shear rate. This approach can be applied to platelet adhesion research.