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Scleral fibroblasts of the chick embryo differentiate into chondrocytes in soft-agar culture

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Summary

Scleral fibroblasts, perichondrial cells of the scleral layer of the 12-day chick embryo, always manifest a fibroblastic morphology in monolayer culture. In soft-agar culture, these cells produce two types of colonies. One type of colony, F-type, consists of adherent fibroblastic cells, and the other, C-type, is composed of scattered round chondrocytic cells. Cells of the C-type colony are surrounded by a halo of extracellular matrix, positive with Alcian blue and with an antibody to cartilage-specific proteoglycan. When a single fibroblast clone in monolayer, derived from a single scleral fibroblast, is subcultured into soft agar, the cells give rise to both C-type and F-type colonies. Further, it was found that cells constituting F-type colonies eventually separate and become spherical, and the F-type colony converts to a C-type colony (C-type conversion). In regard to the C-type convertibility, the primary fibroblast clones were divided into four categories, early time differentiating, middle-time differentiating, late-time differentiating and nondifferentiating. This suggests that the scleral perichondrial layer of the 12-day chick embryo is composed of a variety of cells with different chondrogenic potentialities maintained in each individual cell.

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References

  1. Amprino, R.; Bairati, A. Studi sulle transformazioni delle cartilagini del l’uomo nell’accrescimento e nella senes cenza. I. Cartilagini jaline. Z. Zellfor. Microsk. Anat. 20:143–205; 1934.

    Article  Google Scholar 

  2. Atsumi, T.; Miwa, Y.; Kimata, K., et al. A chondrogenic cell line derived from a differentiating culture of AT805 teratocarcinoma cells. Cell Differ. Dev. 30:109–116; 1990.

    Article  PubMed  CAS  Google Scholar 

  3. 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 

  4. Fujioka, M.; Shimamoto, N.; Kawahara, A., et al. Purification of an autocrine growth factor in conditioned medium obtained from primary cultures of scleral fibroblasts of the chick embryo. Exp. Cell Res. 181:400–408; 1989.

    Article  PubMed  CAS  Google Scholar 

  5. Hall, B. K. Scleral cartilage. In: Hall, B., ed. Developmental and cellular skeletal biology. New York: Academic Press; 1978:46–49.

    Google Scholar 

  6. Hamburger, V.; Hamilton, H. A series of normal stages in the development of the chick. J. Morphol. 88:49–92; 1951.

    Article  Google Scholar 

  7. Horwitz, A. L.; Dorfman, A. The growth of the cartilage cells in soft agar and liquid suspension. J. Cell Biol. 45:434–437; 1970.

    Article  PubMed  CAS  Google Scholar 

  8. Kato, Y.; Iwamoto, M.; Tatsuya, K. Fibroblast growth factor stimulates colony formation of differentiated chondrocytes in soft agar. J. Cell. Physiol. 133:491–498; 1987.

    Article  PubMed  CAS  Google Scholar 

  9. Kimata, K.; Oike, Y.; Tani, K., et al. A large chondroitin sulphate proteoglycan (PG-M) synthesized before chondrogenesis in the limb bud of chick embryo. J. Biol. Chem. 261:13517–13525; 1986.

    PubMed  CAS  Google Scholar 

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

    PubMed  CAS  Google Scholar 

  11. Newsome, D. A.In vitro stimulation of cartilage in embryonic chick neural crest cells by products of retinal pigmented epithelium. Dev. Biol. 49:496–507; 1976.

    Article  PubMed  CAS  Google Scholar 

  12. Pechak, D. G.; Kujawa, M. J.; Caplan, A. I. Morphological and histochemical events during first bone formation in embryonic chick limb. Bone 7:441–458; 1986.

    Article  PubMed  CAS  Google Scholar 

  13. Reinbold, R. Role du tapetum dans la differentiation de la sclerotique chez l’embryon de poulet. J. Embryol. Exp. Morphol. 19:43–47; 1968.

    PubMed  CAS  Google Scholar 

  14. Shinomura, T.; Kimata, K.; Oike, Y., et al. Appearance of distinct types of proteoglycan in a well-defined temporal and spatial pattern during early cartilage formation in the chick limb. Dev. Biol. 103:211–220; 1984.

    Article  PubMed  CAS  Google Scholar 

  15. Shinomura, T.; Jensen, K. L.; Yamagata, M., et al. The distribution of mesenchyme proteoglycan (PG-M) during wing bud outgrowth. Anat. Embryol. 181:227–233; 1990.

    Article  PubMed  CAS  Google Scholar 

  16. Shinomura, T.; Kimata, K. Precartilage condensation during skeletal pattern formation. Dev. Growth & Differ. 32:243–248; 1990.

    Article  Google Scholar 

  17. Smith, L.; Thorogood, P. Transfilter studies on the mechanism of epithelial-mesenchymal interactions leading to chondrogenic differentiation of neural crest cells. J. Embryol. Exp. Morphol. 75:165–188; 1983.

    PubMed  CAS  Google Scholar 

  18. Solursh, M.; Linsenmayer, T. F.; Jensen, K. L. Chondrogenesis from single limb bud mesenchyme cells. Dev. Biol. 94:259–264; 1982.

    Article  PubMed  CAS  Google Scholar 

  19. Stewart, P. A.; McCallion, D. J. Establishment of scleral cartilage in the chick. Dev. Biol. 46:383–389; 1975.

    Article  PubMed  CAS  Google Scholar 

  20. Watanabe, K.; Fujioka, M.; Takeshita, T., et al. Scleral fibroblasts of the chick embryo proliferate by an autocrine mechanism in proteinfree primary cultures: differential secretion of growth factors depending on the growth state. Exp. Cell Res. 182:321–329; 1989.

    Article  PubMed  CAS  Google Scholar 

  21. von der Mark, H.; von der Mark, K.; Gay, S. Study of differential collagen synthesis during development of the chick embryo by immunofluorescence. 1. Preparation of collagen type I and type II specific antibodies and their application to early stages of the chick embryo. Dev. Biol. 48:237–249; 1976.

    Article  PubMed  Google Scholar 

  22. Watt, F. M.; Dudhia, J. Prolonged expression of differentiated phenotype by chondrocytes cultured at low density on a composite substrate of collagen and agarose that restricts cell spreading. Differentiation 38:140–147; 1988.

    Article  PubMed  CAS  Google Scholar 

  23. Wolpert, L.; Stein, W. D. Positional information and pattern formation. In: Malacinski, G. M.; Bryant, S. V., eds. Pattern formation. Macmillan; 1984:3–21.

    Google Scholar 

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Watanabe, K., Yagi, K., Ohya, Y. et al. Scleral fibroblasts of the chick embryo differentiate into chondrocytes in soft-agar culture. In Vitro Cell Dev Biol - Animal 28, 603–608 (1992). https://doi.org/10.1007/BF02631034

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  • DOI: https://doi.org/10.1007/BF02631034

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