Skip to main content
SpringerLink
Log in

Monocyte-macrophage differentiation in vitro: Modulation by extracellular matrix protein substratum

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

The influence of extracellular matrix (ECM) on monocyte-macrophage (mo-mφ) differentiation was investigated using an in vitro model with human peripheral blood mononuclear cells (PBMC) maintained on different matrix protein substrata. Macrophage specific markers associated with differentiation studied were, (a) endocytosis of modified proteins; (b) appearance of mφ specific matrix metalloproteinases (MMPs); (c) activities of myeloperoxidase (MPO) and β-D-glucuronidase; (d) changes in the expression of cell surface antigens. As the duration of monocytes in culture increased, a progressive increase in the rate of differentiation was seen as evidenced by mφ specific functions such as endocytosis of 125[I] acetyl BSA and the appearance of gelatinases A and B. Significantly higher rate of endocytosis and production of MMPs were found in monocytes maintained on fibronectin (FN) and COL I than on COL IV (FN > COL I > COL IV) indicating that cells in contact with stromal components differentiate at a faster rate. FACS analysis done on cells maintained in vitro for phenotypic profile characteristic to mo-mφ differentiation showed downregulation of CD14 occurring in a substratum dependent manner viz, (FN > COL IV > COL I) and upregulation of CD71 was high in cells maintained on COL I and COL IV. Intracellular enzymatic activities such as MPO significantly decreased irrespective of matrix substrata, while β-D-glucuronidase activity increased in a substratum dependent manner (FN > COL I > COL IV). Pretreatment of cells with genistein significantly decreased the secretion of MMPs, particularly MMP 9 in cells maintained on ECM protein (FN) indicating a phosphorylation dependent signalling process in mediating matrix effect. The results of these in vitro studies on mφ specific markers suggest that apart from the diffusible factors, the microenvironment as provided by various matrix proteins particularly FN can modulate mo-mφ differentiation.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Butcher EC: Leukocyte-endothelial cell recognition. Three (or more) steps to specificity and diversity. Cell 67: 1033–1036, 1991

    Google Scholar 

  2. Butcher EC: Lymphocyte homing and homeostasis. Science 272: 60–66, 1996

    Google Scholar 

  3. Dunon D, Piali L, Imhoff BA: To stick or not to stick: The new leukocyte homing paradigm. Curr Opin Cell Biol 8: 714–723, 1996

    Google Scholar 

  4. Ley K, Gaetgens P, Fennie C, Singer MS, Lasky LA, Rosen SD: Lectin like cell adhesion molecule I mediates leukocyte rolling in mesentric venules in vivo. Blood 77: 2553–2560, 1991

    Google Scholar 

  5. von Andrian VH, Chambers JD, McEvoy LM, Bargatse RF, Arfors KE, Butcher EC: Two step model of leukocyte-endothelial interaction in inflammation: Distinct roles for LECAM 1 and the leukocyte αβ integrins in vivo. Proc Natl Acad Sci USA 88: 7538–7545, 1991

    Google Scholar 

  6. Stacker SA, Springer TA: Leukocyte integrin p150, 95 (CD11c/CD18) functions as an adhesion molecule binding to a counter receptor on stimulated endothelium. J Immunol 146: 648–655, 1991

    Google Scholar 

  7. Kodama T, Freeman M, Rohrer L, Zabrecky D, Matsudaira P, Kreiger M: Type I macrophage scavenger receptor contains α-helical and collagen like coiled coils. Nature 343: 531–535, 1990

    Google Scholar 

  8. Schwartz CJ, Valente AJ, Sprague EA, Kelley JL, Nerem RM: The pathogenesis of atherosclerosis. An overview. Clin Cardiol 14(suppl 1): 11–16, 1991

    Google Scholar 

  9. Ross R: The pathogenesis of atherosclerosis: A perspective for 1990s. Nature 362: 801–809, 1993

    Google Scholar 

  10. Rusten LA, Smeland EB, Jacobsen FW, Lieu E, Lesslauer W, Loetscher H, Dubois C M, Jacobsen SE: Tumor necrosis factor alpha inhibits stem cell factor-induced proliferation of human bone marrow progenitor cells in vitro. Role of p55 and p75 tumor necrosis factor receptors. J Clin Invest 94: 165–172, 1993

    Google Scholar 

  11. Shah PK, Falk E, Badimon JJ, Fernandez-Oritz A, Mailhac A, Villareal Levy G, Fallon JT, Regnstrom J, Fuster V: Human monocyte derived macrophages induce collagen breakdown in fibrous caps of atheroscle17 rotic plaques: Potential role of matrix degrading metalloproteinases and implications for plaque rupture. Circulation 92: 1565–1569, 1995

    Google Scholar 

  12. Galis ZS, Sukhova GK, Lark MW, Libby P: Increased expression of matrix metalloproteinases and matrix degrading activity in vulnerable regions of human atherosclerotic plaques. J Clin Invest 94: 2493–2503, 1994

    Google Scholar 

  13. Lin CQ, Bissel MJ: Multifaceted regulation of cell differentiation by ECM. FASEB J 7: 737–743, 1993

    Google Scholar 

  14. Ben Zeev A, Robinson GS, Butcher NLR, Farmer SR: Cell-cell and cell-matrix interaction differentially regulate the expression of hepatic and cytoskeletal genes in primary culture of rat hepatocytes. Proc Natl Acad Sci 85: 2161–2165, 1988

    Google Scholar 

  15. Sudhakaran PR, Stamatoglou SC, Hughes RC: Modulation of protein synthesis and secretion by substratum in primary cultures of rat hepatocytes. Exp Cell Res 167: 505–513, 1986

    Google Scholar 

  16. Hynes RO: Integrins: Versatality, modulation and signalling in cell adhesion. Cell 69: 11–25, 1992

    Google Scholar 

  17. Engvall E, Ruoslahti E: Binding of soluble form of fibroblast surface protein, fibronectin to collagen. Int J Cancer 20: 1–6, 1977

    Google Scholar 

  18. Huh HY, Pearce SF, Yesner LM, Schindler JL, Silverstein RL: Regulated expression of CD36 in foam cell formation. Blood 87: 2020–2025, 1996

    Google Scholar 

  19. Basu SK, Goldstein JL, Anderson RGW, Brown MS: Degradation of cationised LDL and regulation of cholesterol metabolism in homozygous familial hypercholesterolemia fibroblasts. Proc Natl Acad Sci USA 73: 3178–3184, 1976

    Google Scholar 

  20. Greenwood FC, Hunter WM, Clover JS: The preparation of 131[I] labelled human growth hormone of high specific reactivity. Biochem J 89: 114–122, 1963

    Google Scholar 

  21. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ: Protein measurement with folin-phenol reagent. J Biol Chem 193: 265–275, 1951

    Google Scholar 

  22. Ambili M, Sudhakaran PR: Modulation of neutral matrix metalloproteinases of involuting rat mammary gland by different cations and glycosaminoglycans. J Cell Biochem 73: 218–226, 1999

    Google Scholar 

  23. Desser RK, Himmelhoch SR, Evans WH, Januska M, Mage M, Shelton E: Guinea pig heterophil and neutrophil peroxidase. Arch Biochem Biophys 148: 452–465, 1972

    Google Scholar 

  24. Kawai Y, Anno K: Mucopolysaccharide-degrading enzymes from the liver of the squid, Ommastrephes sloani pacifus. I. Hyaluronidase. Biochem Biophys Acta 242: 428–434, 1971

    Google Scholar 

  25. Provvedini DM, Deftos LJ, Manolagas SC: 1,25-Dihydroxyvitamin D3 promotes in vitro morpholegic and enzymatic changes in normal human monocytes consistent with their differentiation into macrophages. Bone 7: 23–28, 1971

    Google Scholar 

  26. Schwartz CJ, Valente AJ, Sprague EA: A modern view of atherogenesis. Am J Cardiol 71: 9B–14B, 1993

    Google Scholar 

  27. Pearson AM: Scavenger receptors in innate immunity. Curr Opin Immunol 8: 20–28, 1996

    Google Scholar 

  28. Rigotti A, Acton SL, Kreiger M: The class B scavenger receptors SR B1 and CD36 are receptors for anionic phospholipids. J Biol Chem 270: 16221–16225, 1995

    Google Scholar 

  29. Bu G, Rennke S: Receptor associated protein is a folding chaperone for LDL receptor-related protein. J Biol Chem 271: 22218–22224, 1996

    Google Scholar 

  30. Kodama T, Reddy P, Kishomoto C, Kreiger M: Purification and characterisation of bovine acetyl LDL receptor. Proc Natl Acad Sci USA 85: 9238–9242, 1996

    Google Scholar 

  31. Via DP, Pons L, Dennison DK, Fanslow AE, Berini F: Induction of acetyl LDL receptor activity by phorbol ester in human monocytic cell line THP 1. J Lipid Res 30: 1515–1524, 1989

    Google Scholar 

  32. Leppert D, Waubant E, Galardy R, Bunnet NW, Hauser SL: T Cell gelatinases mediate in vitro basement membrane transmigration. J Immunol 154: 4379–4386, 1995

    Google Scholar 

  33. Nagase H, Woessner JF: Matrix metalloproteinases. J Biol Chem 274: 151–160, 1999

    Google Scholar 

  34. Esparza J, Vilardel C, Calvo J, Juan M, Vives J, Urbano-Marquez A, Yague J, Cid CM: Fibronectin upregulates Gelatinase B (MMP-9) and induces coordinated expression of Gelatinase A (MMP-2) and its activator MT1-MMP (MMP-14) by human T lymphocyte cell line. A process repressed through RAS/MAP kinase signalling pathways. Blood 94: 2754–2766, 1999

    Google Scholar 

  35. Xei B, Laouar A, Huberman E: Fibronectin mediated cell adhesion is required for the induction of 92 kDa type IV collagenase/Gelatinase (MMP-9) gene expression during macrophage differentiation. J Biol Chem 273: 11576–11582, 1998

    Google Scholar 

  36. Wesley RB II, Meng X, Godin D, Galis SZ: Extracellular matrix modulates macrophage functions characteristic to atheroma. Arterioscler Thromb Vasc Biol 18: 432–440, 1998

    Google Scholar 

  37. Wright SD, Ramos RA, Tobias PS, Ulevitch RJ, Mathison JC: CD14, a receptor for complexes of LPS and LPS binding protein. Science 249: 1431–1433, 1990

    Google Scholar 

  38. Eischen A, Vincent F, Louis B, Schmitt-Goguel M, Bohbot A, Bergerat JP, Oberling F: Immunophenotypic characterisation of human peritoneal and alveolar macrophages and of human blood monocytes differentiated in the presence of either GM-CSF or M-CSF or a combination of GM-CSF/M-CSF. Nouv Rev Fr Hematol 34: 421–434, 1992

    Google Scholar 

  39. Andreesen R, Brugger W, Scheibenbogen C, Kreutz M, Leser HG, Rehm A, Lohr GW: Surface phenotype analysis of human monocyte to macrophage maturation. J Leukocyt Biol 47: 490–497, 1990

    Google Scholar 

  40. McCullough KC, Schaffner R, Kihm Y, Natale VAI, Summerfield A: The phenotype of porcine monocytic cells: Modulation of surface molecules expression upon monocyte differentiation into macrophages. Vet Immunol Immunopathol 58: 265–275, 1997

    Google Scholar 

  41. Shapira L, Soskolne WA, Van Dyke TE: Prostaglandin E2 secretion, cell maturation, and CD14 expression by monocyte-derived macrophages from localised juvenile periodontitis patients. J Periodontol 67: 224–228, 1996

    Google Scholar 

  42. Juliano RL, Haskill S: Signal transduction from the extracellular matrix. J Cell Biol 120: 577–585, 1993

    Google Scholar 

  43. Lin TH, Rosales C, Mondal K, Bolen JB, Haskill S, Juliano RL: Integrin mediated tyrosine phosphorylation and cytokine message induction in monocytic cells: A possible signalling role for the Syk tyrosine kinase. J Biol Chem 270: 16189–16199, 1995

    Google Scholar 

  44. Laouar A, Collart FR, Chubb CBH, Xei B, Huberman E: Interaction between α5β1 integrin and secreted fibronectin is involved in macrophage differentiation of human HL-60 myeloid leukemia cells. J Immunol 162: 407–414, 1999

    Google Scholar 

  45. Hay ED: Collagen and other matrix glycoproteins in embryogenesis. In: E.D. Hay (ed). Cell Biology of Extracellular Matrix. Plenum Press, New York, p 419, 1991

    Google Scholar 

  46. Hay ED: Extracellular matrix alters epithelial differentiation. Curr Opin Cell Biol 5: 1029–1036, 1993

    Google Scholar 

  47. Wright TN: The vascular extracellular matrix. In: V. Fuster, R. Ross, E. Topol (eds). Atherosclerosis and Coronary Artery Disease. Raven Press Ltd., New York, p 4, 1995

    Google Scholar 

Download references

Authors
  1. Shiney Susan Jacob
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Padma Shastry
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. P.R. Sudhakaran
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Jacob, S.S., Shastry, P. & Sudhakaran, P. Monocyte-macrophage differentiation in vitro: Modulation by extracellular matrix protein substratum. Mol Cell Biochem 233, 9–17 (2002). https://doi.org/10.1023/A:1015593232347

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1015593232347

Search

Navigation