Flow Cytometric Analysis of Immune Cells Within Murine Aorta

Methods Mol Biol. 2015;1339:161-75. doi: 10.1007/978-1-4939-2929-0_11.

Abstract

The immune system plays a critical role in the modulation of atherogenesis at all stages of the disease. However, there are many technical difficulties when studying the immune system within murine aortas. Common techniques such as PCR and immunohistochemistry have answered many questions about the presence of immune cells and mediators of inflammation within the aorta yet many questions remain unanswered due to the limitations of these techniques. On the other hand, cumulatively the flow cytometry approach has propelled the immunology field forward but it has been challenging to apply this technique to aortic tissues. Here, we describe the methodology to isolate and characterize the immune cells within the murine aorta and provide examples of functional assays for aortic leukocytes using flow cytometry. The method involves the harvesting and enzymatic digestion of the aorta, extracellular and intracellular protein staining, and a subsequent flow cytometric analysis.

Keywords: Atherosclerosis; Flow cytometry; Leukocytes; Mouse aorta.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Aorta / immunology*
  • Aorta / metabolism
  • Aorta / pathology
  • Aortic Diseases / immunology*
  • Aortic Diseases / metabolism
  • Aortic Diseases / pathology
  • Atherosclerosis / immunology*
  • Atherosclerosis / metabolism
  • Atherosclerosis / pathology
  • Biomarkers / metabolism
  • Cell Separation / methods*
  • Disease Models, Animal
  • Flow Cytometry*
  • Immunophenotyping
  • Leukocytes / immunology*
  • Leukocytes / metabolism
  • Leukocytes / pathology
  • Mice
  • Phenotype

Substances

  • Biomarkers