Skip to main content
SpringerLink
Log in

Autonomous migration of human fetal skin fibroblasts into a denuded area in a cell monolayer is mediated by basic fibroblast growth factor and collagen

  • Growth, Differentiation, And Senescence
  • Published:
In Vitro Cellular & Developmental Biology - Animal Aims and scope Submit manuscript

Summary

Human fetal skin fibroblasts (TIG-3S) were found to migrate into a denuded area in a cell monolayer when cultured in both serum-depleted and serum-supplemented media, unlike adult-donor skin fibroblasts which migrated well only when cultured in serum-supplemented medium. Therefore, a series of experiments was carried out to determine whether autocrine factors are involved in their migration. The migration of TIG-3S cells in serum-depleted medium was suppressed by the addition of suramin, a factor with growth factor antagonist properties, which suggests that growth factors are important for cell migration. The suramin-induced inhibition was reversed completely by adding excess basic fibroblast growth factor (bFGF) to the culture medium and partially by platelet-derived growth factor (PDGF). Treatment with neutralizing anti-PDGF antibody did not suppress TIG-3S cell migration, whereas neutralizing anti-bFGF antibody did, which indicates that bFGF is an autocrine and PDGF a paracrine factor involved in cell migration. Next, an experiment was performed to ascertain whether the extracellular matrix is involved in TIG-3S cell migration. Monensin, an inhibitor of extracellular matrix secretion, inhibited cell migration, which was reversed by adding excess type I collagen, but not excess plasma fibronectin. In addition, further evidence for the involvement of collagen was provided by the observation that ethyl-3,4-dihydroxybenzoate, a specific inhibitor of collagen synthesis, suppressed cell migration. These results suggest that the autonomous migration of TIG-3S human fetal skin fibroblasts is mediated by bFGF and type I collagen, which they produce and secrete.

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. Augustin-Voss, H. G.; Pauli, U. B. Quantitative analysis of autocrine-regulated, matrix-induced, and tumor cell-stimulated endothelial cell migration using a silicon template compartmentalization technique. Exp. Cell Res. 198:221–227; 1992

    Article  PubMed  CAS  Google Scholar 

  2. Betholez, C.; Westermark, E. R. B.; Heldin, C.-E. Coexpression of a PDGF-like growth factor and PDGF receptors in a human oesteosarcoma cell line: implications for autocrine receptor activator. Cell 39:447–457; 1984.

    Article  Google Scholar 

  3. Boyden, S. V. The chemotactic effect of mixtures of antibody and antigen on polymorphonuclear leukocytes. J. Exp. Med. 115:453–466; 1962.

    Article  PubMed  CAS  Google Scholar 

  4. Clemmons, D. R. Age dependent production of a competence factor by human fibroblasts. J. Cell. Physiol. 114:61–67; 1983.

    Article  PubMed  CAS  Google Scholar 

  5. Coffey, B. J., Jr.; Leof, E. B.; Shipley, G. D., et al. Nature of the interaction of growth factor with suramin. J. Cell. Physiol. 132:143–148; 1987.

    Article  PubMed  CAS  Google Scholar 

  6. Flaumenhaft, R.; Moscatelli, D.; Saksela, O., et al. Role of extracellular matrix in the action of basic fibroblast growth factor: matrix as a source of growth factor for long-term stimulation of plasminogen activator production and DNA synthesis. J. Cell. Physiol. 140:75–81; 1989.

    Article  PubMed  CAS  Google Scholar 

  7. Gauss-Muller, V.; Kleinman, H. K.; Martin, G. R., et al. Role of attachment factors and attractants in fibroblast chemotaxisis. J. Lab. Clin. Med. 96:1071–1080; 1980.

    PubMed  CAS  Google Scholar 

  8. Grey, A.-M.; Schor, A. M.; Rushton, G., et al. Purification of the migration stimulating factor produced by fetal and breast cancer patient fibroblasts. Proc. Natl. Acad. Sci. USA 86:2438–2442; 1989.

    Article  PubMed  CAS  Google Scholar 

  9. Kojima, S.; Harpel, P. C.; Rifkin, D. B. Lipoprotein (a) inhibits the generation of transforming growth factor-β: an endogenous inhibitor of smooth muscle cell migration. J. Cell Biol. 113:1439–1445; 1991.

    Article  PubMed  CAS  Google Scholar 

  10. Kondo, H.; Nomaguchi, T. A.; Sakurai, Y., et al. Effects of serum from human subjects of various ages on proliferation of human lung and skin fibroblasts. Exp. Cell Res. 178:287–295; 1988.

    Article  PubMed  CAS  Google Scholar 

  11. Kondo, H.; Nomaguchi, T. A.; Yonezawa, Y. Effects of serum from human subjects of different ages on migration in vitro of human fibroblasts. Mech. Ageing Dev. 47:25–37; 1989.

    Article  PubMed  CAS  Google Scholar 

  12. Kondo, H.; Yonezawa, Y.; Nomaguchi, T. A. Difference in migratory ability between human lung and skin fibroblasts. Zool. Sci. 7:39–46; 1990.

    Google Scholar 

  13. Kondo, H.; Yonezawa, Y. Changes in the migratory ability of human lung and skin fibroblasts during in vitro aging and in vivo cellular senescence. Mech. Ageing Dev. 63:223–233; 1992.

    Article  PubMed  CAS  Google Scholar 

  14. Kondo, H.; Matsuda, R.; Yonezawa, Y. Platelet-derived growth factor in combination with collagen promotes the migration of human skin fibroblasts into a denuded area of a cell monolayer. Exp. Cell Res. 202:45–51; 1992.

    Article  PubMed  CAS  Google Scholar 

  15. Martin, G. R. Laminin and other basement membrane components. Ann. Rev. Cell Biol. 3:57–85; 1987.

    PubMed  CAS  Google Scholar 

  16. Matsuzaki, K.; Yoshitake, Y.; Matuo, Y., et. al. Monoclonal antibodies against heparin-binding growth factor II/basic fibroblast growth factor that block its biological activity. Proc. Natl. Acad. Sci. USA 86:9911–9915; 1989.

    Article  PubMed  CAS  Google Scholar 

  17. Middaugh, C. R.; Mach, H.; Burke, C. J., et al. Nature of the interaction of growth factor with suramin. Biochemistry 31:9016–9024; 1992.

    Article  PubMed  CAS  Google Scholar 

  18. Mignatti, P.; Morimoto, T.; Rifkin, D. B. Basic fibroblast growth factor released by single, isolated cells stimulates their migration on an autocrine manner. Proc. Natl. Acad. Sci. USA 88:11007–11011; 1991.

    Article  PubMed  CAS  Google Scholar 

  19. Minniti, C. P.; Maggi, M.; Helman, L. J. Suramin inhibits the growth of human Rhodomyosarcoma by interrupting the insulin like growth factor autocrine growth loop. Cancer Res. 52:1830–1835; 1992.

    PubMed  CAS  Google Scholar 

  20. Mundy, G. R.; De Martino, S.; Rowe, D. W. Collagen and collagen-derived fragments are chemotactic for tumor cells. J. Clin. Invest. 68:1102–1105; 1981.

    PubMed  CAS  Google Scholar 

  21. Nandan, D.; Clarke, E. P.; Ball, E. H., et al. Ethyl-3,4-dihydroxybenzoate inhibits myoblast differentiation: evidence for an essential role of collagens. J. Cell Biol. 110:1673–1679; 1990.

    Article  PubMed  CAS  Google Scholar 

  22. Pienta, K. J.; Isaacs, W. B.; Vindvich, D., et al. The effects of basic fibroblast growth factor and suramin on cell motility and growth of rat prostate cancer cells. J. Urol. 145:199–202; 1991.

    PubMed  CAS  Google Scholar 

  23. Postlethwaite, A. E.; Seyer, J. M.; Kang, A. H. Chemotactic attraction of human fibroblasts to type I, II and III collagens and collagen-derived peptides. Proc. Natl. Acad. Sci. USA 75:871–875; 1978.

    Article  PubMed  CAS  Google Scholar 

  24. Postlethwaite, A. E.; Keski-Oja, J.; Moses, H. L., et al. Stimulation of the chemotactic migration of human fibroblasts by transforming growth factor-β. J. Exp. Med. 165:251–256; 1987.

    Article  PubMed  CAS  Google Scholar 

  25. Presta, M.; Maier, J. A. M.; Rusnati, M., et al. Basic fibroblast growth factor is released from endothelial extracellular matrix in a biologically active form. J. Cell. Physiol. 140:68–74; 1989.

    Article  PubMed  CAS  Google Scholar 

  26. Root, L. L.; Shipley, G. D. Human dermal fibroblasts express multiple bFGF and aFGF proteins. In Vitro Cell. Dev. Bio. 27A:815–822; 1991.

    CAS  Google Scholar 

  27. Rosen, E. M.; Goldberg, I. D. Protein factors which regulate cell motility. In Vitro Cell. Dev. Biol. 25:1079–1087; 1989.

    Article  PubMed  CAS  Google Scholar 

  28. Ruoslahti, E.; Pierschbacher, M. D. New perspectives in cell adhesion: RGD and integrins. Science 238:491–497; 1987.

    Article  PubMed  CAS  Google Scholar 

  29. Saksela, O.; Rifkin, D. B. Release of basic fibroblast growth factor-heparan sulfate complexes from endothelial cells by plasminogen activator-mediated proteolytic activity. J. Cell Biol. 110:767–775; 1990.

    Article  PubMed  CAS  Google Scholar 

  30. Sasaki, T.; Majamaa, K.; Uitto, J. Reduction of collagen production in Keloid fibroblast cultures by ethyl-3,4-dihydroxybenzoate. J. Biol. Chem. 262:9397–9403; 1987.

    PubMed  CAS  Google Scholar 

  31. Sato, Y.; Rifkin, D. B. Autocrine activities of basic fibroblast growth factor: regulation of endothelial cell movement, plasminogen activator synthesis, and DNA synthesis. J. Cell Biol. 107:1199–1205; 1988.

    Article  PubMed  CAS  Google Scholar 

  32. Sato, Y.; Tsuboi, R.; Lyons, R., et al. Characterization of the activation of latent TGF-β by co-cultures of endothelial cells and pericytes or smooth muscle cells: a self regulating system. J. Cell Biol. 111:757–763; 1990.

    Article  PubMed  CAS  Google Scholar 

  33. Sato, Y.; Hamanaka, R.; Ono, J., et al. The stimulatory effect of PDGF on vascular smooth muscle cell migration is mediated by the induction of endogenous basic FGF. Biochem. Biophys. Res. Commun. 174:1260–1266; 1991.

    Article  PubMed  CAS  Google Scholar 

  34. Schor, S. L.; Schor, A. M.; Rushton, G., et al. Adult, foetal and transformed fibroblasts display different migratory phenotypes on collagen gels: evidence for an isoformic transition during foetal development. J. Cell Sci. 73:221–234; 1985.

    PubMed  CAS  Google Scholar 

  35. Schor, S. L.; Schor, A. M.; Grey, A. M., et al. Foetal and cancer patient fibroblasts produce an autocrine migration-stimulating factor not made by normal adult cells. J. Cell Sci. 90:391–399; 1988.

    PubMed  CAS  Google Scholar 

  36. Senior, R. M.; Griffin, G. L.; Mecham, R. P. Chemotactic responses of fibroblasts to tropoelastin and elastin-derived peptides. J. Clin. Invest. 70:614–618; 1982.

    Article  PubMed  CAS  Google Scholar 

  37. Senior, R. M.; Huang, S. S.; Griffin, G. L., et al. Brain-derived growth factor is a chemoattractant for fibroblasts and astroglial cells. Biochem. Biophys. Res. Commun. 141:67–72; 1986.

    Article  PubMed  CAS  Google Scholar 

  38. Seppa, H.; Grotendorst, G.; Seppa, S., et al. Platelet-derived growth factor is chemotactic for fibroblasts. J. Cell Biol. 92:584–588; 1982.

    Article  PubMed  CAS  Google Scholar 

  39. Stenn, K. S. Quantitative assay of dissociated tissue cell motility in vitro. In Vitro 16:357–360; 1980.

    PubMed  CAS  Google Scholar 

  40. Stoker, M.; Gherardi, E. Regulation of cell movement: The motogenic cytokines. Biochim. Biophys. Acta 1072:81–102; 1991.

    PubMed  CAS  Google Scholar 

  41. Uchida, N.; Smilowitz, H.; Tanzer, M. L. Monovalent ionophores inhibit secretion of procollagen and fibronectin from cultured human fibroblasts. Proc. Natl. Acad. Sci. USA 76:1868–1872; 1979.

    Article  PubMed  CAS  Google Scholar 

  42. Yonezawa, Y.; Kondo, H.; Kaji, K., et al. Species-specific differences in the mitogenic activity of heparin-binding growth factors in the sera of various mammals. Experientia 48:239–245; 1992.

    Article  PubMed  CAS  Google Scholar 

  43. Yoshida, T.; Kawase, M.; Sasaki, K., et al. Comparative studies on the sensitivity of three methods for detecting mycoplasmal contamination in cell cultures. Bull. Jpn. Fed. Cul. Coll. 4:9–15; 1988.

    Google Scholar 

  44. Wharton, W. Newborn human skin fibroblasts senescence in vitro without acquiring adult growth factor requirements. Exp. Cell Res. 154:311–314; 1984.

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Experimental Biology, Tokyo Metropolitan Institute of Gerontology, 35-2 Sakae, Itabashi-ku, 173, Tokyo, Japan

    Hiroshi Kondo, Rei Matsuda & Yumiko Yonezawa

Authors
  1. Hiroshi Kondo
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Rei Matsuda
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yumiko Yonezawa
    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

Kondo, H., Matsuda, R. & Yonezawa, Y. Autonomous migration of human fetal skin fibroblasts into a denuded area in a cell monolayer is mediated by basic fibroblast growth factor and collagen. In Vitro Cell Dev Biol - Animal 29, 929–935 (1993). https://doi.org/10.1007/BF02634231

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02634231

Key words

Search

Navigation