VEGF and ECGF Induce Directed Migration of Endothelial Cells: Qualitative and Quantitative Assay

Proliferation of blood vessels is a process necessary for the normal growth and development of tissues, and for body's repair. Several components are involved in the formation of new blood vessels: chemotactic activity which causes endothelial cells to migrate from existing capillaries into the growing zone; mitogenic activity, which includes mitoses in the endothelial cells; and the organization of the cells to form an organ the new capillary. Several factors were found which induce angiogenesis. Some of them were isolated from malignant tissues and others from normal tissues. We developed a sensitive method of endothelial cell migration towards angiogenic factors. The factors which were chosen: VEGF (Vascular Endothelial Growth Factor), and ECGF (Endothelial Cell Growth Factor). The method is based on migration of cells ABAE (Aortic Arch Derived Bovine Endothelial Cells) under agarose, towards angiogenic factors, which were seeded in wells drilled in the agarose. A dose response curve for the two compounds studied, was found. The optimal migration for VEGF was 10 μg/ml, and for ECGF 100 ng/ml. Our method can be used as a sensitive and accurate system to identify potential angiogenic factors, and to measure their effect on endothelial cell migration.