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Isolation and Characterization of Aortic Dendritic Cells and Lymphocytes in Atherosclerosis.

Methods in molecular biology (Clifton, N.J.) (2017-01-08)
Tae Jin Yun, Jun Seong Lee, Dahee Shim, Jae-Hoon Choi, Cheolho Cheong
RESUMEN

Dendritic cells (DCs) are central to initiate antigen-specific immunity and tolerance. The in vivo development and distribution of DCs are now better understood even in nonlymphoid tissues [1]. Atherosclerosis is a chronic inflammatory disease of blood vessels and DCs are highly enriched in the intimal area of the aorta, which is predisposed to develop atherosclerosis. Previously, we were the first to show antigen presenting DCs and their subsets in the aorta [2, 3]. Here, we discuss several useful methods to characterize not only DCs but also other immune cells in steady state and atherosclerotic aorta. These comprise multiparameter flow cytometry strategies including intracellular staining and cell sorting, en face immunohistochemistry of DCs and regulatory T cells (Tregs), and Oil Red O staining of atherosclerotic lesions in the aorta.

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Roche
DNasa I, from bovine pancreas
Sigma-Aldrich
Colagenasa from Clostridium histolyticum, for general use, Type I, ≥125 CDU/mg solid
Sigma-Aldrich
Colagenasa from Clostridium histolyticum, Type XI, 2-5 FALGPA units/mg solid, ≥800 CDU/mg solid
Hyaluronidase, European Pharmacopoeia (EP) Reference Standard