Abstract
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.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Chesney, J. and Bucala, R. (2001) Peripheral Blood Fibrocytes. In: In Human Cell Culture, Vol. 5. B. O. Pallsson, M. R. Koller, and J. Masters, eds. Kluwer Academic Publishers, London, p. 209–219.
Bucala, R., Spiegel, L. A., Chesney, J., Hogan, M., and Cerami, A. (1994) Circulating fibrocytes define a new leukocyte subpopulation that mediates tissue repair. Mol Med 1, 71–81.
Paget, J. (1863) Lectures on Surgical Pathology. In Royal College of Surgeons of England. Longmans, London.
Stirling, G. A. and Kakkar, V. V. (1969) Cells in the circulating blood capable of producing connective tissue. Br. J. Exp. Pathol. 50, 51–55.
Burkitt, Y. and Heath, H. (1997) Wheater’s Functional Histology, Vol. 1. Churchill Livingston, London, p. 404.
Dunphy, J. (1963) The Fibroblast-A unique Ally for the Surgeon. New Engl. J. Med. 268, 1367–1377.
Brown, J., Greaves, M. F., and Molgaard., H. V. (1991) The gene encoding the stem cell antigen, CD34, is conserved in mouse and expressed in haemopoietic progenitor cell lines, brain, and embryonic fibroblasts. Int. Immunol. 3, 175–184.
Fina, L., Molgaard, H. V., Robertson, D., et al. (1990) Expression of the CD34 gene in vascular endothelial cells. Blood 75, 2417–2426.
Barth, P. J., Ebrahimsade, S., Ramaswamy, A., and Moll, R. (2002) CD34+ fibrocytes in invasive ductal carcinoma, ductal carcinoma in situ, and benign breast lesions. Virchows Arch. 440, 298–303.
Hirohata, S., Yanagida, T., Nagai, T., et al. (2001) Induction of fibroblast-like cells from CD34(+) progenitor cells of the bone marrow in rheumatoid arthritis. J. Leukoc. Biol. 70, 413–421.
Barth, P. J., Ebrahimsade, S., Hellinger, A., Moll, R., and Ramaswamy, A. (2002) CD34+ fibrocytes in neoplastic and inflammatory pancreatic lesions. Virchows Arch. 440, 128–133.
Aiba, S. and Tagami, H. (1997) Inverse correlation between CD34 expression and proline-4-hydroxylase immunoreactivity on spindle cells noted in hypertrophic scars and keloids. J. Cutan. Pathol. 24, 65–69.
Aiba, S., Tabata, N., Ohtani, H., and Tagami, H. (1994) CD34+ spindle-shaped cells selectively disappear from the skin lesion of scleroderma. Arch. Dermatol. 130, 593–597.
Schmidt, M., Sun, G., Stacey, M. A., Mori, L. and Mattoli, S. (2003) Identification of circulating fibrocytes as precursors of bronchial myofibroblasts in asthma. J. Immunol. 171, 380–389.
Chesney, J., Bacher, M., Bender, A., and Bucala, R. (1997) The peripheral blood fibrocyte is a potent antigen-presenting cell capable of priming naive T cells in situ. Proc. Natl. Acad. Sci. USA 94, 6307–6312.
Kirchmann, T. T., Prieto, V. G., and Smoller, B. R. (1994) CD34 staining pattern distinguishes basal cell carcinoma from trichoepithelioma. Arch. Dermatol. 130, 589–592.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2007 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Quan, T.E., Bucala, R. (2007). Culture and Analysis of Circulating Fibrocytes. In: Cope, A.P. (eds) Arthritis Research. Methods in Molecular Medicine, vol 135. Humana Press. https://doi.org/10.1007/978-1-59745-401-8_28
Download citation
DOI: https://doi.org/10.1007/978-1-59745-401-8_28
Publisher Name: Humana Press
Print ISBN: 978-1-58829-344-2
Online ISBN: 978-1-59745-401-8
eBook Packages: Springer Protocols