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Development of a cell line from skin of goldfish, Carassius auratus, and effects of ascorbic acid on collagen deposition

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Abstract

Growth characteristics and collagen expression were investigated in GFSk-S1, a cell line derived from the skin of an adult goldfish (Carassius auratus). These cells are anchorage dependent, grow well in Leibovitz-15 medium with 10% fetal bovine serum, and have been subcultured routinely for 5 years. Cells at various passages have been successfully cryopreserved and thawed. GFSk-S1 cells show mainly a fibroblastic morphology at low density, but at confluence islands of epithelial-shaped cells appear among the fibroblastic cells. The cells require little maintenance, and cultures have been kept viable for more than 3 months without medium changes. Although best growth was observed at room temperature, cell proliferation still occurred at 28°C, and a subline was maintained and passaged for over a year at 25°C. Cells were exposed to various concentrations of ascorbic acid, and its effects on collagen secretion were monitored by light and electron microscopy. Under phase-contrast microscopy, confluent GFSk-S1 cells exposed to ascorbic acid at 50 μg/ml showed distinct development of fibres as early as 3 days after treatment. Histochemical staining for collagen demonstrated a thick network of fibres under a monolayer of ascorbic acid- treated GFSk-S1 cells, and observation by transmission electron microscopy showed collagen fibres with typical banding pattern. This cell line appears to show a stable genotype, as collagen expression was induced at all passages. GFSk-S1 could be useful for studies not only of regulation of protein synthesis, but also of cell differentiation and wound healing

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References

  • Anderson, C.D. & Roberts, R.J. (1975) A comparison of effects of temperature on wound healing in a tropical and temperate teleost. J. Fish Biol. 7, 173-82.

    Article  Google Scholar 

  • Barnes, M.J. (1975) Function of ascorbic acid in collagen metabolism. Ann. NY Acad. Sci. 258, 264-87.

    Article  PubMed  CAS  Google Scholar 

  • Berman, J., Stoner, G., Dawe, C., Rice, J. & Kingsbury, E. (1978) Histochemical demonstration of collagen fibers in ascorbic acid-fed cell cultures. In Vitro 14, 675-85.

    Article  PubMed  CAS  Google Scholar 

  • Bols, N.C. & Lee, L.E.J. (1991) Technology and uses of cell cultures from the tissues and organs of bony fish. Cytotechnology 6, 163-87.

    Article  Google Scholar 

  • Bols, N.C. & Lee, L.E.J. (1994) Cell lines: availability, propagation and isolation. In Biochemistry and Molecular Biology of Fishes. Vol. 3, Analytical techniques (edited by Hochachka, P.W. & Mommsen, T.P.), pp. 145-59. Amsterdam: Elsevier.

    Google Scholar 

  • Bols, N.C., Mosser, D.D. & Steels, G.B. (1992) Temperature studies and recent advances with fish cells in vitro. Comp. Biochem. Physiol. 103A, 1-14.

    Article  Google Scholar 

  • Bols, N.C., Barlian, A., Chirino-trejo, M., Caldwell, S.J., Goegan, P. & L.E.J. Lee (1994a) Development of a cell line from primary cultures of rainbow trout, Oncorhynchus mykiss (Walbaum), gills. J. Fish Dis. 17, 601-11.

    Article  Google Scholar 

  • Bols, N.C., Ganassin, R.C., Tom, D.J. & Lee, L.E.J. (1994b) Growth of fish cell lines in glutamine-free media. Cytotechnology 16, 159-66.

    Article  PubMed  CAS  Google Scholar 

  • Dabrowski, K. (1990) Gulonolactone oxidase is missing in teleost fish. The direct spectrophotometric assay. Biol. Chem. Hoppe-Seyler 371, 207-14.

    PubMed  CAS  Google Scholar 

  • Davidson, G.A., Ellis, A.E. & Secombes, C.J. (1993) Novel cell types isolated from the skin of rainbow trout, Oncorhynchus mykiss. J. Fish Biol. 42, 301-6.

    Article  Google Scholar 

  • Fryer, J.L. & Lannan, C.N. (1994) Three decades of fish cell culture: A current listing of cell lines derived from fishes. J. Tissue Culture Meth. 16, 87-94.

    Article  Google Scholar 

  • Gallop, P.M. & Seifter, S. (1963) Preparation and properties of soluble collagens. In Methods in Enzymology, Vol. VI (edited by Colowick, S.P. & Kaplan, N.O.), pp. 635-641. New York: Academic Press.

    Chapter  Google Scholar 

  • Gauger, A., Robertson, C., Greenlee, J.R., T.K. & Riederer-Henderson, M.A. (1985) A low serum medium for tendon cells: effects of growth factors on tendon cell growth and collagen production. In Vitro Cell. Dev. Biol. 21, 291-6.

    Article  PubMed  CAS  Google Scholar 

  • Geesin, J.C., Brown, L.J., Gordon, J.S. & Berg, R.A. (1993) Regulation of collagen synthesis in human dermal fibroblasts in contracted gels by ascorbic acid, growth factors and inhibitors of lipid peroxidation. Exp, Cell Res. 206, 283-90.

    Article  CAS  Google Scholar 

  • Gould, B.S. (1968) Treatise on Collagen. Vol. 2. Biology of Collagen. New York: Academic Press.

    Google Scholar 

  • Green, H. & Goldberg, B. (1968) Differentiation for collagen synthesis in cultured cells. In Differentiation and Immunology. Symposia of the International Society for Cell Biology. Vol. 7 (edited by Warren, K.B.), pp. 123-34. New York: Academic Press.

    Google Scholar 

  • Halver, J.E., Smith, R.R., Tolbert, B.M. & Baker, E.M. (1975) Utilization of ascorbic acid in fish. Ann. NY Acad. Sci. 258, 81-102.

    Article  PubMed  CAS  Google Scholar 

  • Hayat, M.A. (1986) Basic Techniques for Transmission Electron Microscopy. New York: Academic Press.

    Google Scholar 

  • Hibiya, T., Yokote, M., Oguri, M., Sato, H., Takashima, F & Aida, K. (1982) An Atlas of Fish Histology: Normal and Pathological Features. Tokyo: Kodansha Ltd.

    Google Scholar 

  • Kaneko, Y., Igarashi, M., Iwashita, M., Suzuki, K., Kojima, H., Kimura, S. & Hasobe, M. (1995) Effects of fish and calf type I collagens as culture substrate in the adhesion and spreading process of established fish cells. In Vitro Cell. Dev. Biol. 31, 178-82.

    Article  CAS  Google Scholar 

  • Kimura, S., Miyauchi, Y. & Uchida, N. (1991) Scale and bone type I collagens of carp (Cyprinus carpio). Comp. Biochem. Physiol. 99B, 473-6.

    CAS  Google Scholar 

  • Kirchhofer, D., Reinhardt, C.A. & Zbinden, G. (1986) Collagen synthesis in growing human skin fibroblasts. Exp. Cell Biol. 54, 177-82.

    Article  PubMed  CAS  Google Scholar 

  • Lee, L.E.J. & Bols, N.C. (1991) Identification of fibronectin in rainbow trout and influence of cortisol on fibronectin accumulation by rainbow trout cell cultures. J. Aquatic Anim. Health 3, 31-40.

    Article  Google Scholar 

  • Lee, L.E.J., Clemons, J.H., Bechtel, D.G., Caldwell, S.J., Han, K.B., Pasitschniakarts, M., Mosser, D.D. & Bols, N.C. (1993) Development and characterization of a rainbow trout liver cell line expressing cytochrome P450-dependent monooxygenase activity. Cell Biol. Toxicol. 9, 279-94.

    Article  PubMed  CAS  Google Scholar 

  • Levene, C.I. & Bates, C.J. (1975) Ascorbic acid and collagen synthesis in cultured fibroblasts. Ann. NY Acad. Sci. 258, 288-306.

    Article  PubMed  CAS  Google Scholar 

  • Lillie, R.D. (1965) Histopathologic Technic and Practical Histochemistry, 3rd edn. pp. 535-6. New York: McGraw-Hill.

    Google Scholar 

  • Lim, C. & Lovell, R.T. (1978) Pathology of the vitamin C deficiency syndrome in channel catfish. J. Nutr. 108, 1137-46.

    PubMed  CAS  Google Scholar 

  • Liu, E., Minor, R.R., Horovitz, O., Wootton, J.A.M., Podleski, T.R. & Salpeter, M.M. (1993) Secreted collagen induced by ascorbic acid in L5 cloned muscle cultures does not affect acetylcholine receptor expression. Exp. Cell Res. 209, 76-81.

    Article  PubMed  CAS  Google Scholar 

  • Marty, G.D. & Summerfelt, R.C. (1990) Wound healing in channel catfish by epithelialization and contraction of granulation tissue. Trans. Am. Fish. Soc. 119, 145-50.

    Article  Google Scholar 

  • Matsumoto, J., Lynch, T.J., Grabowski, S.M., Taylor, J.D. & Tchen, T.T. (1982) Introduction of melanized cells from a goldfish erythrophoroma: isolation of pigment translocation variants. Science 217, 1149-51.

    Article  PubMed  CAS  Google Scholar 

  • Montero, P., Borderias, J., Turnay, J. & Leyzarbe, M.A. (1990) Characterization of hake (Merluccius merluccius L.) and trout (Salmo irideus Gibb) collagen. J. Agric. Food Chem. 38, 604-9.

    Article  CAS  Google Scholar 

  • Mothersill, C., Lyng, F., Lyons, M. & Cottell, D. (1995) Growth and differentiation of epidermal cells from the rainbow trout established as explants and maintained in various media. J. Fish Biol. 46, 1011-25.

    Article  Google Scholar 

  • Murad, S., Grove, D., Linberg, K.A. Reynolds, G., Sivarajah, A. & Pinnell, S.R. (1981) Regulation of collagen synthesis by ascorbic acid. Proc. Natl. Acad. Sci. USA 78, 2879-82.

    Article  PubMed  CAS  Google Scholar 

  • Nimni, M.E. (1988) Collagen, Vol. 1. Florida: CRC Press.

    Google Scholar 

  • Quinones, S.R., Neblock, D.S. & Berg, R.A. (1986) Regulation of collagen production and collagen mRNA amounts in fibroblasts in response to culture conditions. Biochem. J. 239, 179-83.

    PubMed  CAS  Google Scholar 

  • Saito, H., Iwami, H. & Shigeoka, T. (1991) In vitro cytotoxicity of 45 pesticides to goldfish GF-Scale (GFS) cells. Chemosphere 23, 525-537.

    Article  CAS  Google Scholar 

  • Saito, H., Koyasu, J. & Shigeoka, T. (1993) Cytotoxicity of anilines and aldehydes to goldfish GFS cells and relationships with 1-octanol/water partition coefficients. Chemosphere 27, 1553-60.

    Article  CAS  Google Scholar 

  • Sandness, K. (1991) Vitamin C in fish nutrition — a review. Fisk. Dir. Skr. Ser. Ernaering 4, 3-32.

    Google Scholar 

  • Sappino, A.P., Schurch, W. & Gabbiani, G. (1990) Biology of disease. Differentiation repertoire of fibroblastic cells: expression of cytoskeletal proteins as marker of phenotypic modulations. Lab. Invest. 63, 144-61.

    PubMed  CAS  Google Scholar 

  • Sato, M., Yoshinaka, R. & Ikeda, S. (1978) Dietary ascorbic acid requirement of rainbow trout for growth and collagen formation. Bull. Japan. Soc. Sci. Fish. 44, 1029-35.

    CAS  Google Scholar 

  • Yoshinaka, R., Sa, K., Sato, M. & Anbe, H. (199) Distribution of collagen in body of several fishes. Nippon Suisan Gakkaishi 56, 549.

    Google Scholar 

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  1. Department of Veterinary Anatomy, Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, SK, Canada, S7N 5B4

    L. E. J. Lee, S. J. Caldwell & J. Gibbons

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  1. L. E. J. Lee
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  2. S. J. Caldwell
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  3. J. Gibbons
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Lee, L.E.J., Caldwell, S.J. & Gibbons, J. Development of a cell line from skin of goldfish, Carassius auratus, and effects of ascorbic acid on collagen deposition. J Mol Hist 29, 31–43 (1997). https://doi.org/10.1023/A:1026412817431

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