In the cardiovascular system spironolactone and amiloride exert beneficial effects that increase over time. Surface expression of epithelial sodium channels and mechanical cell properties are major determinants of endothelial function and are regulated by these drugs. This, however, has never been studied over a long time period. A major problem is that such experiments performed in standard cell cultures are limited due to potential changes in endothelial phenotype and possible loss of differentiation. To overcome this problem we established a culture model of ex vivo arteries which should allow to study endothelial cells (ECs) closer to natural conditions and thus should permit long-term investigations with more physiological impact. Aortae excised from mice were cleaned from surrounding tissue and cut into small rings. Rings were cut open, glued with Cell-Tak^TM on coverslips with the EC layer facing upwards and cultured for up to three weeks in conventional culture medium. Integrity of the endothelial cell layer and preservation of endothelial identity over time were weekly verified by VE-cadherin immunostaining. As evident from fluorescence microscopy, the EC monolayer of ex vivo arteries was found intact up to at least three weeks. VE-cadherin, outlining the individual cells, was preserved at all times. In contrast to mono-cultured ECs, ECs of the ex vivo arteries were aligned in a regular order reflecting the direction of shear stress and the organisation of underlying tissue.We conclude that ex vivo arteries kept in culture could serve as a suitable model for long-term evaluation of endothelial function.