Differentiation of Human Monocytes In Vitro: A Model of Macrophage Ontogeny

  • R. Andreesen
  • M. Kreutz
Part of the Ernst Schering Research Foundation Workshop book series (SCHERING FOUND, volume 11)


Monocytes are the common “precursor” of the different types of macrophages which are distributed ubiquitous in all tissues. Monocytes and granulocytes both originate from committed progenitor cells in the bone marrow (colony-forming unit granulocyte-macrophage, CFU-GM). Glycoprotein hormones termed colony-stimulating factors [CSF; macrophage CSF (M-CSF), granulocyte-macrophage CSF (GM-CSF)] or interleukin-3 (IL-3; multi-CSF) regulate the differentiation of this stem cell into monoblasts which then differentiate into promonocytes (Metcalf 1991).


Human Monocyte Mononuclear Phagocyte System Monocyte Differentiation Vitronectin Receptor Human Blood Monocyte 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Akiyama Y, Miller PJ, Thurman GB, Neubauer RH, Oliver C, Favilla T, Beman JA, Oldham RK, Stevenson HC (1983) Characterization of a blood monocyte subset with low peroxidase activity. J Clin Invest 72: 1093–1105PubMedCrossRefGoogle Scholar
  2. Akiyama Y, Griffith R, Miller P, Stevenson GW, Lund S, Kanapa DJ, Stevenson HC (1988) Effects of adherence, activation and distinct serum proteins on the in vitro human monocyte maturation process. J Leukoc Biol 43: 224–231PubMedGoogle Scholar
  3. Andreesen R, Picht J, Löhr GW (1983a) Primary cultures of human blood-born macrophages grown on hydrophobic teflon membranes. J Immunol Methods 56: 295–304PubMedCrossRefGoogle Scholar
  4. Andreesen R, Osterholz J, Bross KJ, Schulz A, Löhr GW (1983b) Cytotoxic effector cell function at different stages of human monocyte-macrophage maturation. Cancer Res 43: 5931–5936PubMedGoogle Scholar
  5. Andreesen R, Osterholz J, Bross KJ, Costabel U, Löhr GW (1984) Expression of transferrin receptors and intracellular ferritin during terminal differentiation of human monocytes. Blut 49: 195–202PubMedCrossRefGoogle Scholar
  6. Andreesen R, Bross KJ, Osterholz J, Emmrich F (1986) Human macrophage maturation and heterogeneity: analysis with a newly generated set of monoclonal antibodies to differentiation antigens. Blood 67: 1257–1264PubMedGoogle Scholar
  7. Andreesen R, Gadd S, Costabel U, Leser HG, Speth V, Cesnik B, Atkins RC (1988a) Human macrophage maturation and heterogeneity: restricted expression of late differentiation antigens in situ. Cell Tissue Res 253: 271–279PubMedCrossRefGoogle Scholar
  8. Andreesen R, Gadd S, Brugger W, Löhr GW, Atkins RC (1988b) Activation of human monocyte-derived macrophages cultured on Teflon: response to interferon-gamma during terminal maturation in vitro. Immunobiology 177: 186–198PubMedCrossRefGoogle Scholar
  9. Andreesen R, Brugger W, Scheibenbogen C, Kreutz M, Leser HG, Rehm A, Löhr GW (1990a) Surface phenotype analysis of human monocyte to macrophage maturation. J Leukoc Biol 47: 490–497PubMedGoogle Scholar
  10. Andreesen R, Scheibenbogen C, Kreutz M, Krause S, Rehm A (1990b) Regulation of neopterin secretion during human monocyte to macrophage differentiation in vitro. Pteridines 2: 59–61Google Scholar
  11. Anegon I, Blottiere H, Cuturi MS, Lenne, Trinchieri G, Faust J, Perussia B (1993) Characterization of a human monocyte antigen, B148.4, regulated during cell differentiation and activation. J Leukoc Biol 53: 390–398PubMedGoogle Scholar
  12. Arenson BE, Epstein MB, Seeger RC (1980) Volumetric and functional heterogeneity of human monocytes. J Clin Invest 65: 613–618PubMedCrossRefGoogle Scholar
  13. Bar-Shavit Z, Teitelbaum SL, Reitsma P, Hall A, Pegg LE, Trial J, Kahn AJ (1983) Induction of monocytic differentiation and bone resorption by 1,25dihydroxyvitamin D3. Proc Natl Acad Sci USA 80: 5907–5911PubMedCrossRefGoogle Scholar
  14. Becker S, Warren MK, Haskill S (1987) Colony-stimulating factor-induced monocyte survival and differentiation into macrophages in serum-free culture. J Immunol 139: 3703–3709PubMedGoogle Scholar
  15. Breitmann TR, Selonick SE, Collins SJ (1980) Induction of differentiation of the human promyelocytic leukemia cell line (HL-60) by retinoic acid. Proc Natl Acad Sci USA 77: 2936–2940CrossRefGoogle Scholar
  16. Brugger W, Andreesen R (1991) Inhibition of in vitro differentiation of human monocytes to macrophages by lipopolysaccharide (LPS): phenotypic and functional analysis. Int Immunol 3: 221–227PubMedCrossRefGoogle Scholar
  17. Brugger W, Kreutz M, Andreesen R (1991) Macrophage colony-stimulating factor is required for human monocyte survival and acts as a cofactor for their terminal differentiation to macrophages in vitro. J Leukoc Biol 49: 483–488PubMedGoogle Scholar
  18. Cassileth PA, Suholet D, Cooper RA (1981) Early changes in phosphatidylcholine metabolism in human acute promyelocytic leukemia cells stimulated to differentiate by phorbol ester. Blood 58: 237PubMedGoogle Scholar
  19. Clarkson SB, Ory PA (1988) Developmentally regulated IgG Fc receptors on cultured human monocytes. J Exp Med 167: 408–417PubMedCrossRefGoogle Scholar
  20. Clohisy DR, Bar-Shavit Z, Chappel JC, Teitelbaum S (1987) 1,25-Dihydroxyvitamin D3 modulates bone marrow macrophage precursor proliferation and differentiation. Up-regulation of the mannose receptor. J Biol Chem 262: 15922–15929Google Scholar
  21. Collins SJ, Ruscetti FW, Gallagher RE, Gallo RC (1979) Normal functional characteristics of cultured human promyelocytic leukemia cells (HL-60) after induction of differentiation by dimethylsulfoxide. J Exp Med 149: 969PubMedCrossRefGoogle Scholar
  22. Dodd RC, Cohen MS, Newman SL, Gray TK (1983) Vitamin D metabolites change the phenotype of monoblastic U937 cells. Proc Natl Acad Sci USA 80: 7538–7541PubMedCrossRefGoogle Scholar
  23. Dougerty GJ, McBride WH (1984) Macrophage heterogeneity. J Clin Lab Immunol 14: 1–11Google Scholar
  24. Eischen A, Vincent F, Bergerat JP, Louis B, Faradji A, Bohbot A, Oberling F (1991) Long-term cultures of human monocytes in vitro. Impact of GMCSF on survival and differentiation. J Immunol Methods 143: 209–221Google Scholar
  25. Ezekowitz RAB, Stahl PD (1988) The structure and function of vertebrate mannose lectin-like proteins. J Cell Sci Suppl 9: 212–133Google Scholar
  26. Firestein GS, Zvaifler NJ (1987) Down regulation of human monocyte differentiation antigens by interferon. Cell Immunol 104: 343–354PubMedCrossRefGoogle Scholar
  27. Goerdt S, Bhardway R, Sorg C (1994) Inducible expression of MS-1 high molecular weight protein by endothelial cells of continuous origin and by dendritic cells/macrophages in vivo and in vitro. Am J Pathol (in press)Google Scholar
  28. Harwix S, Andreesen R, Ferber E, Schwamberger G (1992) Human macrophages secrete a tumoricidal activity distinct from tumour necrosis factor-a and reactive nitrogen intermediates. Res Immunol 143: 89–94PubMedCrossRefGoogle Scholar
  29. Haskill S, Johnson C, Eierman D, Becker S, Warren K (1988) Adherence induces selective mRNA expression of monocyte mediators and protooncogenes. J Immunol 140: 1690–1694PubMedGoogle Scholar
  30. Helinski EH, Bielat KL, Ovak GM, Pauly JL (1988) Long-term cultivation of functional human macrophages in teflon dishes with serum-free media. J Leukoc Biol 44: 111–121PubMedGoogle Scholar
  31. Johnston RB, Zucker-Franklin D (1988) The mononuclear phagocyte system. In: Zucker-Franklin D, Greaves MF, Grossi CE, Marmont AM (eds) Atlas of blood cells: function and pathology. Fischer, StuttgartGoogle Scholar
  32. Jungi TW, Hafner S (1986) Quantitative assessment of Fc receptor expression and function during in vitro differentiation of human monocytes to macrophages. Immunology 58: 131–137PubMedGoogle Scholar
  33. Kasinrerk W, Baumruker T, Majdic O, Knapp W, Sockinger H (1993) CD1 molecule expression on human monocytes induced by granulocyte-macrophage colony-stimulating factor. J Immunol 150: 579–589PubMedGoogle Scholar
  34. Krause SW, Kreutz M, Zenke G, Andreesen R (1992) Developmental regulation of granulocyte-macrophage colony-stimulating factor production during monocyte-to-macrophage maturation. Ann Hematol 64: 190–195PubMedCrossRefGoogle Scholar
  35. Kreutz M, Andreesen R (1990) Induction of human monocyte into macro- phage maturation by 1,25-dihydroxyvitamin D3. Blood 76: 2457–2461PubMedGoogle Scholar
  36. Kreutz M, Krause SW, Hennemann B, Rehm A, Andreesen R (1992) Macrophage heterogeneity and differentiation: defined serum-free culture conditions induce different types of macrophages in vitro. Res Immunol 143: 107–115PubMedCrossRefGoogle Scholar
  37. Kreutz M, Andreesen R, Krause SW, Ritz E, Reichel H (1993) 1,25-Dihydroxyvitamin D3 production and vitamin D3 receptor expression are developmentally regulated during differentiation of human monocytes into macrophages. Blood 82: 1300–1307Google Scholar
  38. Krissansen GW, Elliot MJ, Lucas CM, Stomski FC, Berndt MC, Cheresh DA, Lopez AF, Burnes GF (1990) Identification of a novel integrin 13 subunit expressed on cultured monocytes (macrophages). J Biol Chem 265: 823–830PubMedGoogle Scholar
  39. Lastres P, Bellon T, Cabanas C, Sanchez-Madrid F, Acevedo A, Gougos A, Letarte M, Bernabeu C (1992) Regulated expression on human macrophages of endoglin, an Arg-Gly-Asp-containing surface antigen. Eur J Immunol 22: 393–397PubMedCrossRefGoogle Scholar
  40. Liu M, Wu M (1992) Induction of human monocyte cell line U937 differentia- tion and CSF-1 production by phorbol ester. Exp Hematol 20: 974–979PubMedGoogle Scholar
  41. Lopez M, Martinache C, Canepa S, Chokri M, Scotto F, Bartholeyns J (1993) Autologous lymphocytes prevent the death of monocytes in culture and promote as do GM-CSF, IL-3 and M-CSF, their differentiation into macrophages. J Immunol Methods 159: 29–38PubMedCrossRefGoogle Scholar
  42. Mantovani A, Bottazzi B, Colotta F, Sozzani S, Ruco L (1992) The origin and function of tumor-associated macrophages. Immunol Today 13: 265–270PubMedCrossRefGoogle Scholar
  43. McKenzie ANJ, Culpepper JA, de Waal Malefyt R, Briere F, Punnonen J, Aversa G, Sato A, Dang W, Cocks BG, Menon S, de Vries JE, Banchereau J, Zurawski G (1993) Interleukin 13, a T-cell-derived cytokine that regulates human monocyte and B-cell function. Proc Natl Acad Sci USA 90: 3735–3739PubMedCrossRefGoogle Scholar
  44. Metcalf D (1991) Control of granulocytes and macrophages: molecular, cellular and clinical aspects. Science 254: 529–533PubMedCrossRefGoogle Scholar
  45. Munn DH, Cheung NV (1989) Antibody-dependent antitumor cytotoxicity by human monocytes cultured with recombinant macrophage colony-stimulating factor. J Exp Med 170: 511–526PubMedCrossRefGoogle Scholar
  46. Munn DH, Cheung NV (1990) Phagocytosis of tumor cells by human monocytes cultured in recombinant macrophage colony-stimulating factor. J Exp Med 172: 231–237PubMedCrossRefGoogle Scholar
  47. Musson RA (1983) Human serum induces maturation of human monocytes in vitro. Changes in cytolytic activity, intracellular lysosomal enzymes, and nonspecific esterase activity. Am J Physiol 111: 331–340Google Scholar
  48. Musson RA, Shafran H, Henson PM (1980) Intracellular levels and stimulated release of lysosomal enzymes from human peripheral blood monocytes and monocyte-derived macrophages. J Reticoendothel Soc 28: 249–264Google Scholar
  49. Nathan CF (1987) Secretory products of macrophages. J Clin Invest 79: 319–323PubMedCrossRefGoogle Scholar
  50. Nichilo MO, Burns GF (1993) Granulocyte-macrophage and macrophage colony-stimulating factors differentially regulate av integrin expression on cultured human macrophages. Proc Natl Acad Sci USA 90: 2517–2521PubMedCrossRefGoogle Scholar
  51. Nichols KE, Weinberg JB (1989) Essential amino acid deprivation induces monocytic differentiation of the human HL-60 myeloid leukemia cell line. Blood 73: 1298–1306PubMedGoogle Scholar
  52. Passlick B, Flieger D, Ziegler-Heitbrock HWL (1989) Identification and characterization of a novel monocyte subpopulation in human peripheral blood. Blood 74: 2527–2534PubMedGoogle Scholar
  53. Peters JH, Ruhl S, Friedrichs D (1987) Veiled accessory cells deduced from monocytes. Immunobiology 176: 154–166PubMedCrossRefGoogle Scholar
  54. Provvedini DM, Deftos LJ, Manolagas SC (1986) 1,25-Dihydroxyvitamin D3 promotes in vitro morphological and enzymatic changes in normal human monocytes consistent with their differentiation into macrophages. Bone 7: 23–28Google Scholar
  55. Raff HV, Picker LJ, Stobo JD (1980) Macrophage heterogeneity in man. A subpopulation of HLA-DR bearing macrophages required for antigen-induced T cell activation also contains stimulators for autologous-reactive T cells. J Exp Med 152: 581–593Google Scholar
  56. Ralph P, Harris PE, Punjabi CJ, Welte K, Litcofsky PB, Ho MK, Rubin BY, Moore MAS, Springer TA (1983) Lymphokine inducing “terminal differentiation” of the human monoblast leukemia line U937: a role for y-interferon. Blood 62: 1169–1175PubMedGoogle Scholar
  57. Rutherford MS, Witsell A, Schook LB (1993) Mechanisms generating functionally heterogeneous macrophages:chaos revisited. J Leukoc Biol 53: 602–618PubMedGoogle Scholar
  58. Ruppert J, Peters JH (1991) IL-6 and IL-1 enhance the accessory activity of human blood monocytes during differentiation to macrophages. J Immunol 146: 144–149PubMedGoogle Scholar
  59. Scheibenbogen C, Andreesen R (1991) Developmental regulation of the cytokine repertoire in human macrophages: IL-1, IL-6, TNF-alpha and M-CSF. J Leukoc Biol 50: 35–42Google Scholar
  60. Scheibenbogen C, Zenke G, Faggs B, Motoyoshi K, Kanz L, Sawert H, Brugger W, Andreesen R (1990) Secretion of colony-stimulating factor for macrophages (M-CSF) and granulocyte/macrophages (GM-CSF) is developmentally regulated in human macrophages. In: Freund M, Link H, Welte K (eds) Cytokines in heopoisis, oncology and aids. Springer, Berlin Heidelberg New York, pp 45–51CrossRefGoogle Scholar
  61. Scheibenbogen C, Moser H, Krause SW, Andreesen R (1992) Interferon. gamma-induced expression of tissue factor activity during human monocyte to macrophage maturation. Haemostasis 22: 173–178PubMedGoogle Scholar
  62. Schlesier M, Krause S, Dräger R, Wolff-Vorbeck G, Kreutz M, Andreesen R (1992) Differential response of the autoreactive T-cell clone UA-S2 to different autologous stimulator cells. International congress of immunology, Budapest, p 336 (W56–42, abstract)Google Scholar
  63. Schreiber RS, Blum JS, Stenson WF, MacDermott RP, Stahl PD, Teitelbaum SL, Perkins SL (1991) Monomeric IgG2a promotes maturation of bone-marrow macrophages and expression of the mannose receptor. Proc Nati Acad Sci USA 88: 1616–1620CrossRefGoogle Scholar
  64. Schumann RR, van der Bosch J, Rüller S, Ernst M, Schlaak M (1989) Monocyte long-term cultivation on microvascular endothelial cell monolayers: morphological and phenotypic characterization and comparison with monocytes cultured on tissue culture plastic. Blood 73: 818–826PubMedGoogle Scholar
  65. Suzu S, Yokota H, Yamada M, Yanai N, Saito M, Kawashima T, Saito M, Takaku F, Motoyoshi K (1989) Enhancing effect of human monocytic colony-stimulating factor on monocyte tumoricidal activity. Cancer Res 49: 5913–5917PubMedGoogle Scholar
  66. Te Velde AA, Klomp JPG, Yard BA, de Vries JE, Figdor CG (1988) Modulation of phenotypic and functional properties of human peripheral blood monocytes by IL-4. J Immunol 140: 1548–1553Google Scholar
  67. Trinchieri G, Rosen M, Perussia B (1987) Retinoic acid cooperates with tumor necrosis factor and immune interferon in inducing differentiation and growth inhibition of the human promyelocytic leukemic cell line HL-60. Blood 69: 1218–1224PubMedGoogle Scholar
  68. Unanue ER, Allen PM (1987) The basis for the immunoregulatory role of macrophages and other accessory cells. Science 236: 551–557PubMedCrossRefGoogle Scholar
  69. Vincent F, Eischen A, Bergerat JP, Faradji A, Bohbot A, Oberling F (1992) Human blood-derived macrophages: differentiation in vitro of large quantity of cells in serum-free medium. Exp Hematol 20: 17–23PubMedGoogle Scholar
  70. Vogel SN, Perera PY, Hogan MM, Majde JA (1988) Use of serum-free, compositionally defined medium for analysis of macrophage differentiation in vitro. J Leukoc Biol 44: 136–142PubMedGoogle Scholar
  71. Von Briesen H, Sander U, Seack KH, Kreutz M, Becker K, Müller S, Rübsamen-Waigmann H, Andreesen R (1992) HIV-induced syncytia formation in human monocytes/macrophages: a microcinematographic study. Pathobiology 60 [Suppl 11: 1–38Google Scholar
  72. Wang SY, Mak KL, Chen LY, Chou MP, Ho CK (1992) Heterogeneity of human blood monocyte: two subpopulations with different sizes, phenotypes and functions. Immunology 77: 298–303PubMedGoogle Scholar
  73. Weinberg JB, Larrick JW (1987) Receptor-mediated monocytoid differentiation of human promyelocytic cells by tumor necrosis factor: synergistic actions with interferon and 1,25-dihydroxyvitamin D3. Blood 70: 994–1002PubMedGoogle Scholar
  74. Wewers MD, Herzyk DJ (1989) Alveolar macrophages differ from blood monocytes in human IL-lbeta release. J Immunol 143: 1635–1641PubMedGoogle Scholar
  75. Yamauchi K, Martinet Y, Crystal RG (1987) Modulation of fibronectin gene expression in human mononuclear phagocytes. J Clin Invest 80: 1720–1727PubMedCrossRefGoogle Scholar
  76. Young DA, Lowe LD, Clark SC (1990) Comparison of the effect of IL-3, granulocyte-macrophage colony-stimulating factor and macrophage colony-stimulating factor in supporting monocyte differentiation in culture. Analysis of antibody-dependent cytotoxicity. J Immunol 145: 607–615Google Scholar
  77. Zaiss M, Krause SW, Kreutz M, Andreesen R (1991) Lymphocyte-mediated inhibition of human monocyte to macrophage maturation in vitro. Proceedings of 5th Annual Conference of the Upper Rhine Universities on the Macrophage, Strasbourg/France, abstract 100Google Scholar
  78. Zembala M, Uracz W, Ruggiero I, Mytar B, Pryjma J (1984) Isolation and functional characteristics of FcR+ and FcR human monocyte subsets. J Immunol 133: 1293–1299PubMedGoogle Scholar
  79. Ziegler-Heitbrock HWL, Ströbel M, Kieper D, Fingerle G, Schlunck T, Peters-mann I, Ellwart J, Blumenstein M, Haas JG (1992) Differential expression of cytokines in human blood monocyte subpopulations. Blood 79: 503–511PubMedGoogle Scholar
  80. Zwadlo G, Bröcker E-B, von Bassewitz D-B, Feige U, Sorg C (1985) A monoclonal antibody to a differentiation antigen present on mature human macrophages and absent from monocytes. J Immunol 134: 1487–1492PubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1994

Authors and Affiliations

  • R. Andreesen
  • M. Kreutz

There are no affiliations available

Personalised recommendations