MO396: Development, Establishment and Validation of in Vitro and EX Vivo Assays of Vascular Calcification

Abstract

Abstract BACKGROUND AND AIMS Vascular calcification is one major complication in patients with chronic kidney disease (CKD), with a misbalance in calcium and phosphate metabolism playing a crucial role. The mechanisms underlying vascular calcification have not been entirely revealed to date. As studies aiming at the identification and characterization of the involved mediators are highly relevant, standardized operating protocols (SOP) for reliable assays to make different studies comparable, are needed. METHOD We are dealing with the following in vitro and ex vivo experimental conditions to study vascular calcifications: induction of calcification by phosphate and CaCl2, incubation time, as well as foetal calve serum concentration and ageing. Furthermore, the location and inhibition of vascular calcifications are studied. Human aortic smooth muscle cells are used for in vitro experiments and rat aorta for ex vivo experiments. The degree of calcification is estimated by quantification of calcium concentrations and by von Kossa staining. RESULTS As a result, a step-by-step protocol for performing experiments on vascular calcification was created. CONCLUSION Through the established protocol and examination of decent modifications nuances, we aim to promote standardized research on vascular calcification.