Liberation and Phenotyping of monocytic cells in human venous tissues
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Abstract
Varicose veins (varices) are a common, progressive degenerative disease of the superficial venous system. Previous histological studies revealed infiltrating immune cells including dendritic cells, macrophages and T cells (Buján et al., 2008, Cherian et al., 1999, Chu et al., 2013, Sayer and Smith, 2004). In the present study, the composition of monocytic cells in varicose veins was analyzed using flow cytometry. For this purpose, a step-by-step mechanical cleansing/tissue digestion process was developed to generate undamaged single cells. Large saphenous veins, removed for bypass surgery, served as non-inflamed control tissue. Flow cytometric analysis of the immune cell infiltrate in varicose veins and bypass veins showed that varicose veins contained up to ten times more CD45+ immune cells than bypass veins and that the immune infiltrate consisted predominantly of monocytic cells. While the outer connective tissue mainly contained CD206 neg myeloid cells, deeper layers of the vein, which became only accessible through digestion, harbored CD206 pos myeloid cells. These observations suggest that CD206 neg monocytes migrate via the connective tissue and differentiate into CD206 pos macrophages in the vein wall.