Flow Cytometry Analysis of Immune Cells Within Murine Aortas

The al goal of the following experiment is to characterize the composition and phenotype of aortic and adventitional infiltrating leukocytes by flow cytometry. This is achieved by first isolating an atherosclerotic aorta as a second step. The surrounding adventitia is isolated from the aorta and single cell suspensions are prepared from both tissues.

Next, the single cell suspensions of fluorescently labeled to differentiate specific subsets of leukocytes from the advent tissue and aortic wall results are obtained that show the diversity of aortic infiltrating leukocytes during atherogenesis and the feasibility of isolating the aortic advent tissue leukocytes based on flow cytometric analysis of single cell suspensions. The main advantage of this technique over existing methods like immunohistochemistry is that leukocytes can be examined and phenotyped using multiple markers from a single aorta. This method can help answer key questions concerning the cellular mediators of inflammation in the pathogenesis of atherosclerosis and other vascular inflammatory disorders.