Technical Advance: The rat aorta contains resident mononuclear phagocytes with proliferative capacity and proangiogenic properties

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Abstract
Methods to target/isolate aorta resident immunocytes and study their angiogenic behavior. Angiogenesis in the aortic ring model is preceded by activation of the immune system and impaired by ablation of adventitial macrophages. Treatment of aortic cultures with M-CSF induced extensive periaortic outgrowth of CD45+ CD68+ mononuclear cells with ultrastructural features of macrophages and DCs. Periaortic lysis of collagen caused many CD45+ CD68+ cells to attach to the bottom of the culture dish. Lifting the collagen gels left behind patches of CD45+ CD68+ cells, which focally organized into branching cords. These cells also expressed CD14, CD169, F4/80, and α-SMA but not CD31, vWF, desmin, or CD163. DNA synthesis studies showed that M-CSF-stimulated cells were actively proliferating. Aortic patch cells showed phagocytic properties and responded to IL-4 and GM-CSF by expressing MHC II, differentiating into DCs, and forming multinucleated giant cells. They also stimulated angiogenesis and VEGF production in aortic ring cultures. This study demonstrates that the rat aorta contains a distinct subset of immature immunocytes capable of proliferating, differentiating into macrophages and DCs, and stimulating angiogenesis. Isolation of these cells in patches from M-CSF-stimulated aortic rings provides a reproducible system to study the biology and angiogenic role of the resident immune system of the aortic wall.
Funding Information
  • National Heart Lung
  • Blood Institute (HL-52585)
  • Merit Review
  • Department of Veterans Affairs Medical Research Services

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