Novel Experimental Approach to Investigate Immune Control of Vascular Function: Co-culture of Murine Aortas With T Lymphocytes or Macrophages.

Abstract

Cardiovascular disease, the current leading cause of death worldwide, is a multifactorial disorder that involves a strong contribution of both the innate and adaptive immune systems. Overactivation of the immune system and inappropriate secretion of pro-inflammatory cytokines lead to vascular impairments and the development of cardiovascular disorders, including hypertension, atherosclerosis, and peripheral artery disease. Lymphocytes, macrophages, and dendritic cells can all secrete pro-inflammatory cytokines. This makes it challenging to isolate a specific subset of immune cells, particularly cytokines, and their contribution to vascular dysfunction remains difficult to elucidate. To solve this problem, our laboratory has developed the novel "immune cell-aorta" co-culture system described herein. This experimental protocol enables investigators to isolate an immune cell of interest and identify the cytokine(s) at the origin of vascular alterations. Key features • Novel ex vivo approach combining the culture of one population of immune cells with blood vessels. • No direct contact between the cells and the blood vessels; the model enables studying the role of immune cell-derived factors or cytokines on vascular function. • Blood vessels can subsequently be used for functional (wire/pressure myography), molecular (western blot, quantitative real-time RT-PCR), and histological studies.