Culture and Analysis of Circulating Fibrocytes

Fibrocytes circulate in the peripheral blood, produce collagen and other matrix proteins, and express cell surface markers indicative of a hematopoetic origin distinguishing them from fibroblasts. Circulating fibrocytes were first identified in 1994 in a model system of wound repair, and defined by their growth characteristics and unique surface phenotype. The methods currently employed for the isolation, growth, and characterization of peripheral blood fibrocytes rely on the entry of “fibroblast-like” cells into wound chambers, or the derivation of “fibroblast-like” cells from the buffy coat of peripheral blood obtained from different mammalian species. In this protocol, we culture fibrocytes from the mononuclear cells of peripheral blood and harvest the cultured cells for flow cytometry analysis.