Skip to main content
SpringerLink
Log in

Restoration of the differentiated functions of serially passaged chondrocytes using staurosporine

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

Summary

Among the various directions explored in order to have a large number of differentiated articular chondrocytes easily available, the restoration of the differentiated properties after cell multiplication in monolayer has been proposed. It has been clearly shown that the synthesis of cartilage proteoglycans and type II collagen synthesis is coincident with the presence of a faint microfibrillar architecture but is absent in chondrocytes showing well-defined actin cables. Staurosporin, mainly described as a protein kinase C inhibitor, has also been shown to rapidly induce the disruption of the actin microfilaments. The purpose of this paper was to investigate whether properties of differentiated chondrocytes were reinitiated upon staurosporin treatment of serially passaged chondrocytes. Results showed, after staurosporine treatment of cells at Passage two for 5 d, complete suppression of type I and type III collagen synthesis and induction of type II collagen synthesis and of Alcian blue stainable matrix. Additionally, we showed that staurosporin restored metabolic responses that chondrocytes in primary culture exhibit upon interleukin-1β treatment (decrease of Alcian blue- positive cells, induction of expression of the 92 kDa gelatinase, nitric oxide production). We conclude that staurosporin is a potent redifferentiating agent of articular chondrocytes that have been subcultured up to Passage two for multiplication. Taking into account that the cellularity of cartilage is very low, staurosporine-treated chondrocytes could be useful as an alternative cellular model to evaluate pharmacotoxicological effects of drugs.

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. Arner, E. C.; Tortorella, M. D. Signal transduction through chondrocyte integrin receptors induces matrix metalloproteinase synthesis and synergizes with interleukin-1. Arthritis Rheum. 38:1304–1314; 1995.

    Article  PubMed  CAS  Google Scholar 

  2. Aydelotte, M. B.; Raiss, R. X.; Caterson, B., et al. Influence of interleukin-1 on the morphology and proteoglycan metabolism of cultured bovine articular chondrocytes. Connect. Tissue Res. 28:143–159; 1992.

    PubMed  CAS  Google Scholar 

  3. Benya, P. D.; Brown, P. D.; Padilla, S. R. Microfilament modification by dihydrocytochalasin B causes retinoic acid- modulated chondrocytes to reexpress the differentiated collagen phenotype without a change in shape. J. Cell Biol. 106:161–170; 1988.

    Article  PubMed  CAS  Google Scholar 

  4. Benya, P. D.; Padilla, S. R. Staurosporine, an inhibitor of protein kinase C, enhances reexpression of the differentiated chondrocyte collagen phenotype: proceedings of the 36th annual meeting of the orthopaedic research society; 1990, February 5–8; New Orleans, Louisiana: 183.

  5. Benya, P. D.; Padilla, S. R. Dihydrocytochalasin B enhances transforming growth factor-β-induced reexpression of the differentiated chondrocyte phenotype without stimulation of collagen synthesis. Exp. Cell Res. 204:268–277; 1993.

    Article  PubMed  CAS  Google Scholar 

  6. Benya, P. D.; Padilla, S. R.; Nimni, M. E. The progeny of rabbit articular chondrocytes synthesize collagen types I and III and type I trimer, but not type II. Verification by cyanogen bromide peptide analysis. Biochemistry 16:865–872; 1977.

    Article  PubMed  CAS  Google Scholar 

  7. Benya, P. D.; Padilla, S. R.; Nimni, M. E. Independent regulation of collagen types by chondrocytes during the loss of differentiated function in culture. Cell 15:1313–1321; 1978.

    Article  PubMed  CAS  Google Scholar 

  8. Benya, P. D.; Shaffer, J. D. Dedifferentiated chondrocytes reexpress the differentiated collagen phenotype when cultured in agarose gels. Cell 30:215–224; 1982.

    Article  PubMed  CAS  Google Scholar 

  9. Bonaventure, J.; Kadhom, N.; Cohen-Solal, L., et al. Reexpression of cartilage-specific genes by dedifferentiated human articular chondrocytes cultured in alginate beads. Exp. Cell Res. 212:97–104; 1994.

    Article  PubMed  CAS  Google Scholar 

  10. Borge, L.; Demignot, S.; Adolphe, M. Type II transglutaminase expression in rabbit articular chondrocytes in culture: relation with cell differentiation, cell growth, cell adhesion and cell apoptosis. Biochim. Biophys. Acta 1312:117–124; 1996.

    Article  PubMed  Google Scholar 

  11. Demignot, S.; Borge, L.; Adolphe, M. Transglutaminase activity in rabbit articular chondrocytes in culture. Biochim. Biophys. Acta 1266:163–170; 1995.

    Article  PubMed  Google Scholar 

  12. Finer, M. H.; Gerstenfeld, L. C.; Young, D., et al. Collagen expression in embryonic chicken chondrocytes treated with phorbol myristate acetate. Mol. Cell. Biol. 5:1415–1424; 1985.

    PubMed  CAS  Google Scholar 

  13. Geng, Y.; Maier, R.; Lotz, M. Tyrosine kinases are involved with the expression of inducible nitric oxide synthase in human articular chondrocytes. J. Cell. Physiol. 163:545–554; 1995.

    Article  PubMed  CAS  Google Scholar 

  14. Green, W. T. Behavior of articular chondrocytes in culture. Clin. Orthop. 75:248–260; 1971.

    Article  PubMed  Google Scholar 

  15. Green, L. C.; Wagner, D. A.; Glogowski, J., et al. Analysis of nitrate, nitrite, and [15N]nitrate in biological fluids. Anal. Biochem. 126:131–138; 1982.

    Article  PubMed  CAS  Google Scholar 

  16. Ito, A.; Nose, T.; Takahashi, S., et al. Cyclooxygenase inhibitors augment the production of pro-matrix metalloproteinase 9 (progelatinase B) in rabbit articular chondrocytes. FEBS Lett. 360:75–79; 1995.

    Article  PubMed  CAS  Google Scholar 

  17. Lev, R.; Spicer, S. S. Specific staining of sulfate groups with Alcian blue at low pH. J. Histochem. Cytochem. 12:309; 1964.

    PubMed  CAS  Google Scholar 

  18. Mallein-Gerin, F.; Garrone, R.; van der Rest, M. Proteoglycan and collagen synthesis are correlated with actin organization in dedifferentiated chondrocytes. Eur. J. Cell Biol. 56:364–373; 1991.

    PubMed  CAS  Google Scholar 

  19. Mallein-Gerin, F.; Ruggiero, F.; Garrone, R. Proteoglycan core protein and type II collagen gene expressions are not correlated with cell shape changes during low density chondrocytes cultures. Differentiation 43:204–211; 1990.

    Article  PubMed  CAS  Google Scholar 

  20. Miossec, P. Pro- and antiinflammatory cytokine balance in rheumatoid arthritis. Clin. Exp. Rheum. 13:S13-S16; 1995.

    Google Scholar 

  21. Mobley, P. L.; Hedberg, K.; Bonin, L., et al. Decreased phosphorylation of four 20-kDa proteins precedes staurosporine- induced disruption of the actin/myosin cytoskeleton in rat astrocytes. Exp. Cell Res. 214:55–66; 1994.

    Article  PubMed  CAS  Google Scholar 

  22. O’Neill, L. A. J. Towards an understanding of the signal transduction pathways for interleukin 1. Biochim. Biophys. Acta 1266:31–44; 1995.

    Article  PubMed  Google Scholar 

  23. Palmer, R. M. J.; Hickery, M. S.; Charles, I. G., et al. Induction of nitric oxide synthase in human chondrocytes. Biochem. Biophys. Res. Commun. 193:398–405; 1993.

    Article  PubMed  CAS  Google Scholar 

  24. Thenet, S.; Benya, P. D.; Demignot, S., et al. SV-40 immortalization of rabbit articular chondrocytes: alteration of differentiated functions. J. Cell. Physiol. 150:158–167; 1992.

    Article  PubMed  CAS  Google Scholar 

  25. von der Mark, K. Differentiation, modulation and dedifferentiation of chondrocytes. Rheumatology 10:272–315; 1986.

    Google Scholar 

  26. Yamamoto, S.; Jiang, H.; Nishikawa, K., et al. Protein kinase C-dependent and -independent actions of a potent protein kinase C inhibitor, staurosporine. Eur. J. Pharmacol. 227:113–122; 1992.

    Article  PubMed  CAS  Google Scholar 

  27. Yu, J. C. M.; Gotlieb, A. I. Disruption of endothelial actin microfilaments by protein kinase C inhibitors. Microvasc. Res. 43:100–111; 1992.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Laboratoire de Pharmacologie Cellulaire de l’Ecole Pratique des Hautes Etudes, Centre de Recherches Biomédicales des Cordeliers, 15 rue de l’Ecole de Médecine, 75006, Paris, France

    Laurence Borge, Francois Lemare, Sylvie Demignot & Monique Adolphe

Authors
  1. Laurence Borge
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Francois Lemare
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Sylvie Demignot
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Monique Adolphe
    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

Borge, L., Lemare, F., Demignot, S. et al. Restoration of the differentiated functions of serially passaged chondrocytes using staurosporine. In Vitro Cell.Dev.Biol.-Animal 33, 703–709 (1997). https://doi.org/10.1007/s11626-997-0128-9

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11626-997-0128-9

Key words

Search

Navigation