Circulating CD45+/Col-1+ fibrocytes are contaminated with polymorphonuclear leukocytes. a Representative gating strategy for identification of circulating CD45+/Collagen-1+ fibrocytes from PBMCs. Isotype control for collagen-1 (Col-1) was used to set the gate for Col-1+ cells within alive CD45+ cells. Red cells are CD45+Col-1+. b FSC and SSC characteristics of CD45+/Col-1+ cells (in red) compared to all alive cells (blue) showing that most CD45+/Col-1+ cells are found in the polymorphonuclear leukocytes fraction. c Histogram overlay showing surface expression of CD15 assessed by flow cytometry on CD45+/Col-1+ cells (red), CD14+ monocytes (black) and T cells (gray). d CD45+/Col-1+ cell enriched fraction and PBMC cultured fibrocytes were analyzed with immunocytochemistry (ICC) for CD15 and collagen-1 expression. Magnification for all ICC figures was 200× and sections were counterstained with hematoxylin. This is representative of 7 experiments
CD14+ Mo = CD14+ monocytes, PBMC = peripheral blood mononuclear cells, FSC = forward scatter, SSC = side scatter.