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. 1982 Oct 1;156(4):1101–1114. doi: 10.1084/jem.156.4.1101

In vitro differentiation of human monocytes. Differences in monocyte phenotypes induced by cultivation on glass or on collagen

PMCID: PMC2186821  PMID: 6961188

Abstract

We demonstrated that the in vitro differentiation of human peripheral blood monocytes to macrophages is dependent on the environment and conditions of monocyte culture. Cultivation of monocytes on glass or microexudate-coated glass gave rise to cells resembling foreign body granuloma macrophages. After an initial rise in Fc receptor- and C3 receptor-mediated phagocytosis, a progressive loss of Fc receptor expression and C3-mediated ingestion were observed. The monocyte surface antigens recognized by the anti-human monocyte monoclonal antibodies 1D5 and 63D3 were lost from the surface of the majority of cells cultured on glass and microexudates. A subpopulation of Fc receptor-positive cells that were 1D5 and 63D3 positive was retained in fully differentiated cell populations. In comparison, monocytes cultivated on collagen matrices gave rise to highly phagocytic cells resembling human resident tissue macrophages. Both Fc- and C3-mediated phagocytosis were enhanced and remained so during the entire length of culture. The surface antigens recognized by the 1D5 antibody, expressed on all freshly seeded monocytes, was maintained on the macrophages. The antigen recognized by the 63D3 antibody was not expressed on mature cells. The present evidence would indicate that variations in expression of phagocytic receptors and the surface antigens 1D5 and 63D3 can be ascribed to the stage of development of the macrophage or its stage of activation, rather than to independent subsets of mononuclear phagocytes.

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Selected References

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  1. Ault K. A., Springer T. A. Cross-reaction of a rat-anti-mouse phagocyte-specific monoclonal antibody (anti-Mac-1) with human monocytes and natural killer cells. J Immunol. 1981 Jan;126(1):359–364. [PubMed] [Google Scholar]
  2. Austyn J. M., Gordon S. F4/80, a monoclonal antibody directed specifically against the mouse macrophage. Eur J Immunol. 1981 Oct;11(10):805–815. doi: 10.1002/eji.1830111013. [DOI] [PubMed] [Google Scholar]
  3. Bennett W. E., Cohn Z. A. The isolation and selected properties of blood monocytes. J Exp Med. 1966 Jan 1;123(1):145–160. doi: 10.1084/jem.123.1.145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Breard J., Reinherz E. L., Kung P. C., Goldstein G., Schlossman S. F. A monoclonal antibody reactive with human peripheral blood monocytes. J Immunol. 1980 Apr;124(4):1943–1948. [PubMed] [Google Scholar]
  5. COHN Z. A., BENSON B. THE DIFFERENTIATION OF MONONUCLEAR PHAGOCYTES. MORPHOLOGY, CYTOCHEMISTRY, AND BIOCHEMISTRY. J Exp Med. 1965 Jan 1;121:153–170. doi: 10.1084/jem.121.1.153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Carrel A., Ebeling A. H. THE FUNDAMENTAL PROPERTIES OF THE FIBROBLAST AND THE MACROPHAGE : II. THE MACROPHAGE. J Exp Med. 1926 Aug 31;44(3):285–305. doi: 10.1084/jem.44.3.285. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chang Y. C., Yao C. S. Investigation of the human macrophage. I. Collection and in vitro cultivation. Eur J Immunol. 1979 Jul;9(7):517–520. doi: 10.1002/eji.1830090706. [DOI] [PubMed] [Google Scholar]
  8. Elsdale T., Bard J. Collagen substrata for studies on cell behavior. J Cell Biol. 1972 Sep;54(3):626–637. doi: 10.1083/jcb.54.3.626. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hirsch S., Austyn J. M., Gordon S. Expression of the macrophage-specific antigen F4/80 during differentiation of mouse bone marrow cells in culture. J Exp Med. 1981 Sep 1;154(3):713–725. doi: 10.1084/jem.154.3.713. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Johnson W. D., Jr, Mei B., Cohn Z. A. The separation, long-term cultivation, and maturation of the human monocyte. J Exp Med. 1977 Dec 1;146(6):1613–1626. doi: 10.1084/jem.146.6.1613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kaplan G. Differences in the mode of phagocytosis with Fc and C3 receptors in macrophages. Scand J Immunol. 1977;6(8):797–807. doi: 10.1111/j.1365-3083.1977.tb02153.x. [DOI] [PubMed] [Google Scholar]
  12. Lewis M. R. The Formation of Macrophages, Epithelioid Cells and Giant Cells from Leucocytes in Incubated Blood. Am J Pathol. 1925 Jan;1(1):91–100.1. [PMC free article] [PubMed] [Google Scholar]
  13. Mariano M., Nikitin T., Malucelli B. E. Immunological and non-immunological phagocytosis by inflammatory macrophages, epithelioid cells and macrophage polykaryons from foreign body granulomata. J Pathol. 1976 Nov;120(3):151–159. doi: 10.1002/path.1711200304. [DOI] [PubMed] [Google Scholar]
  14. Nakagawara A., Nathan C. F., Cohn Z. A. Hydrogen peroxide metabolism in human monocytes during differentiation in vitro. J Clin Invest. 1981 Nov;68(5):1243–1252. doi: 10.1172/JCI110370. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Newman S. L., Musson R. A., Henson P. M. Development of functional complement receptors during in vitro maturation of human monocytes into macrophages. J Immunol. 1980 Nov;125(5):2236–2244. [PubMed] [Google Scholar]
  16. Perussia B., Lebman D., Ip S. H., Rovera G., Trinchieri G. Terminal differentiation surface antigens of myelomonocytic cells are expressed in human promyelocytic leukemia cells (HL60) treated with chemical inducers. Blood. 1981 Oct;58(4):836–843. [PubMed] [Google Scholar]
  17. Postlethwaite A. E., Jackson B. K., Beachey E. H., Kang A. H. Formation of multinucleated giant cells from human monocyte precursors. Mediation by a soluble protein from antigen-and mitogen-stimulated lymphocytes. J Exp Med. 1982 Jan 1;155(1):168–178. doi: 10.1084/jem.155.1.168. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ridley M. J., Ridley D. S., Turk J. L. Surface markers on lymphocytes and cells of the mononuclear phagocyte series in skin sections in leprosy. J Pathol. 1978 Jun;125(2):91–98. doi: 10.1002/path.1711250204. [DOI] [PubMed] [Google Scholar]
  19. Smith B. R., Ault K. A. Increase of surface Ia-like antigen expression on human monocytes independent of antigenic stimuli. J Immunol. 1981 Nov;127(5):2020–2027. [PubMed] [Google Scholar]
  20. Springer T., Galfré G., Secher D. S., Milstein C. Mac-1: a macrophage differentiation antigen identified by monoclonal antibody. Eur J Immunol. 1979 Apr;9(4):301–306. doi: 10.1002/eji.1830090410. [DOI] [PubMed] [Google Scholar]
  21. Todd R. F., 3rd, Griffin J. D., Ritz J., Nadler L. M., Abrams T., Schlossman S. F. Expression of normal monocyte-macrophage differentiation antigens on HL60 promyelocytes undergoing differentiation induced by leukocyte-conditioned medium or phorbol diester. Leuk Res. 1981;5(6):491–495. doi: 10.1016/0145-2126(81)90119-3. [DOI] [PubMed] [Google Scholar]
  22. Ugolini V., Nunez G., Smith R. G., Stastny P., Capra J. D. Initial characterization of monoclonal antibodies against human monocytes. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6764–6768. doi: 10.1073/pnas.77.11.6764. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Wellhausen S. R., Boros D. L. Comparison of Fc, C3 receptors and Ia antigens on the inflammatory macrophage isolated from vigorous or immunomodulated liver granulomas of schistosome-infected mice. J Reticuloendothel Soc. 1981 Sep;30(3):191–203. [PubMed] [Google Scholar]
  24. Werb Z., Bainton D. F., Jones P. A. Degradation of connective tissue matrices by macrophages. III. Morphological and biochemical studies on extracellular, pericellular, and intracellular events in matrix proteolysis by macrophages in culture. J Exp Med. 1980 Dec 1;152(6):1537–1553. doi: 10.1084/jem.152.6.1537. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Werb Z., Banda M. J., Jones P. A. Degradation of connective tissue matrices by macrophages. I. Proteolysis of elastin, glycoproteins, and collagen by proteinases isolated from macrophages. J Exp Med. 1980 Nov 1;152(5):1340–1357. doi: 10.1084/jem.152.5.1340. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Wong L., Wilson J. D. The identification of Fc and C3 receptors on human neutrophils. J Immunol Methods. 1975 Apr;7(1):69–76. doi: 10.1016/0022-1759(75)90131-3. [DOI] [PubMed] [Google Scholar]
  27. Wuest D., Crane R., Rinehart J. J. Enhancement of Fc receptor function during human monocyte differentiation in vitro. J Reticuloendothel Soc. 1981 Aug;30(2):147–155. [PubMed] [Google Scholar]
  28. Yem A. W., Parmely M. J. Modulation of Ia-like antigen expression and antigen-presenting activity of human monocytes by endotoxin and zymosan A. J Immunol. 1981 Dec;127(6):2245–2251. [PubMed] [Google Scholar]
  29. Zola H., McNamara P., Thomas M., Smart I. J., Bradley J. The preparation and properties of monoclonal antibodies against human granulocyte membrane antigens. Br J Haematol. 1981 Jul;48(3):481–490. doi: 10.1111/j.1365-2141.1981.tb02740.x. [DOI] [PubMed] [Google Scholar]
  30. van Furth R. Macrophage activity and clinical immunology. Origin and kinetics of mononuclear phagocytes. Ann N Y Acad Sci. 1976;278:161–175. doi: 10.1111/j.1749-6632.1976.tb47027.x. [DOI] [PubMed] [Google Scholar]

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