Ferric Chloride-induced Murine Thrombosis Models

Wei Li; Marvin Nieman; Anirban Sen Gupta

doi:10.3791/54479

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Method Article

Ferric Chloride-induced Murine Thrombosis Models

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DOI:

10.3791/54479

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September 5th, 2016

Wei Li¹^,² , Marvin Nieman³ , Anirban Sen Gupta⁴

¹Department of Cellular and Molecular Medicine, Lerner Research Institute, Cleveland Clinic, ²Department of Molecular Medicine, Cleveland Clinic Lerner College of Medicine of Case Western Reserve University, ³Department of Pharmacology, Case Western Reserve University, ⁴Department of Biomedical Engineering, Case Western Reserve University

Summary

We report a refined procedure of the ferric chloride (FeCl₃)-induced thrombosis models on carotid and mesenteric artery as well as vein, characterized efficiently using intravital microscopy to monitor time to occlusive thrombi formation.

Abstract

Arterial thrombosis (blood clot) is a common complication of many systemic diseases associated with chronic inflammation, including atherosclerosis, diabetes, obesity, cancer and chronic autoimmune rheumatologic disorders. Thrombi are the cause of most heart attacks, strokes and extremity loss, making thrombosis an extremely important public health problem. Since these thrombi stem from inappropriate platelet activation and subsequent coagulation, targeting these systems therapeutically has important clinical significance for developing safer treatments. Due to the complexities of the hemostatic system, in vitro experiments cannot replicate the blood-to-vessel wall interactions; therefore, in vivo studies are critical to understand pathological mechanisms of thrombus formation. To this end, various thrombosis models have been developed in mice. Among them, ferric chloride (FeCl₃) induced vascular injury is a widely used model of occlusive thrombosis that reports platelet activation and aggregation in the context of an aseptic closed vascular system. This model is based on redox-induced endothelial cell injury, which is simple and sensitive to both anticoagulant and anti-platelets drugs. The time required for the development of a thrombus that occludes blood flow gives a quantitative measure of vascular injury, platelet activation and aggregation that is relevant to thrombotic diseases. We have significantly refined this FeCl₃-induced vascular thrombosis model, which makes the data highly reproducible with minimal variation. Here we describe the model and present representative data from several experimental set-ups that demonstrate the utility of this model in thrombosis research.

Introduction

Arterial thrombosis (blood clot) is a common complication of many systemic diseases associated with chronic inflammation, including atherosclerosis, diabetes, obesity, cancer and chronic autoimmune rheumatologic disorders. Thrombi that occur in the arterial circulation stem from inappropriate platelet activation, aggregation and subsequent coagulatory mechanisms, and are implicated in heart attacks, strokes and extremity loss. The vessel wall is a complex system that includes multiple cell types and is influenced by a multitude of extrinsic factors including shear stress, circulating blood cells, hormones and cytokines, as well as expression of antioxidant proteins in....