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Comparison of trout hepatocyte culture on different substrates

  • Rapid Communications in cell Biology
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Editor's Statement This paper reports improved methods for culture of trout liver-derived cells that make in vitro investigations of fish metabolism, carcinogenesis and chemical toxicity more feasible than previously applied techniques. Recent interest in fish as models for study and indicators of effects of envionmental and food-related toxins make this work timely, poarticularly since many of the compounds of interest are primarily metabolized by hepatocytes or act on liver as a major target. David W. Barnes

Summary

Comparisons were made of attachment and viability of rainbow trout (Salmo gairdneri) hepatocytes in short-term (2 days), primary culture on plastic, collagen-coated or extracellular matrix (ECM) coated dishes. Hepatocyte isolation routinely yielded cells with good viability (96%). Cells plated on ECM attached with high efficiency (93%) in contrast to cells cultured on plastic or collagen (∼20%). The cells plated on ECM flattened out and formed monolayers, while the cells on plastic and collagen rounded up and formed multi-cell aggregates in suspension. Viability of cells in all substrates remained high over the 2 day culture period. ECM is the first substrate to support trout-hepatocyte attachment in primary culture. Differentiated liver function was maintained in cells cultured on ECM as evidence by the induction of tyrosine aminotransferase by hydrocortisone (200%).

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References

  1. Bailey, G. S.; Taylor, M. J.; Selivonchick, D. P. Aflatoxin B1 metabolism and DNA binding in isolated hepatocytes from rainbow trout (Salmo gairdneri). Carcinogenesis 3:511–518; 1982.

    Article  PubMed  CAS  Google Scholar 

  2. Gospodarowicz, D.; Delgado, D.; Vlodavsky, I. Permissive effect of the extracellular matrix on cell proliferationin vitro. Proc. Natl. Acad. Sci. 77:4078–4094; 1980.

    Google Scholar 

  3. Hayashi, S.; Oshiro, Z. Gluconeogenesis and glycolysis in isolated perfused liver of the cel, Bull. Jpn. Soc. Sci. Fish. 41:201–208; 1975.

    CAS  Google Scholar 

  4. Hazel, J. R.; Prosser, C. L. Incorporation of I-14C-Acetate into fatty acids and sterols by isolated hepatocytes of thermally acclimated rainbow trout (Salmo gairdneri). J. Comp. Physiol. 134:321–329; 1979.

    CAS  Google Scholar 

  5. Hendricks, J. D. Chemical carcinogenesis in fish. In: Weber, L. J., ed. Aquatic Toxicology. New York: Raven Press; 1982: 149–211.

    Google Scholar 

  6. Hendricks, J. D.; Meyers, T. R.; Shelton, D. W.; Casteel, J. L.; Bailey, G. S. Hepatocarcinogenicity of benzo[a]pyrene to rainbow trout by dietary exposure and intraperitoneal injection. JNCI 74:839–847; 1985.

    PubMed  CAS  Google Scholar 

  7. Hsu, I. C.; Lipsky, M. M.; Cole, K. E.; Su, C. H.; Trump, B. F. Isolation and culture of hepatocytes from human liver of immediate autopsy. In Vitro Cellular Dev. Biol. 21:154–160; 1985.

    Article  CAS  Google Scholar 

  8. Klaunig, J. E. Establishment of fish hepatocyte cultures for use inin vitro carcinogenicity studies. Natl. Cancer Inst. Mongr. 65:163–173; 1984.

    CAS  Google Scholar 

  9. Klaunig, J. E.; Goldblatt, P. J.; Hinton, D. E.; Lipsky, M. M.; Chako, J.; Trump, B. F. Mouse liver cell culture. II. Primary culture. In Vitro 17:926–934; 1981.

    Article  PubMed  CAS  Google Scholar 

  10. Klaunig, J. E.; Ruch, R. J.; Goldblatt, P. J. Trout hepatocyte culture: isolation and primary culture. In Vitro Cellular Dev. Biol. 21:221–227; 1985.

    Article  CAS  Google Scholar 

  11. Laishes, B. A.; Roberts, E.; Farber, E.In vitro measurement of carcinogen resistant liver cells during hepatocarcinogenesis. Int. J. Cancer. 21:186–193; 1978.

    Article  PubMed  CAS  Google Scholar 

  12. Lin, R. C.; Knox, W. E. Adaptation of the rat liver tyrosine α-ketoglutarate transaminase. Biochem. Biophys. Acta. 26:85–88; 1957.

    Article  PubMed  CAS  Google Scholar 

  13. Michalopoulos, G.; Pitot, H. C. Primary culture of parenchymal liver cells on collagen membranes. Exp. Cell Res. 94:70–78; 1975.

    Article  PubMed  CAS  Google Scholar 

  14. Parker, R. S.; Morrissey, M. T.; Moldeus, P.; Selivonchick, D. P. The use of isolated hepatocytes from rainbow trout (Salmo gardneri) in the metabolism of acetaminophen. Comp. Biochem. Physiol. 70B:631–633; 1981.

    CAS  Google Scholar 

  15. Porthe-Nibelle, J.; Lahlow, B. Mechanisms of glucocorticoid uptake by isolated hepatocytes of the trout. Comp. Biochem. Physiol. 69B:425–433; 1981.

    CAS  Google Scholar 

  16. Saez, L.; Golcoechea, O.; Anthauer, R.; Krauskopf, M. Behavior of RNA and protein synthesis during the acclimatization of the carp. Studies with isolated hepatocytes. Comp. Biochem. Physiol. 72B:31–38; 1982.

    CAS  Google Scholar 

  17. Schoenhard, G. L.; Hendricks, J. D.; Nixon, J. E.; Lee, D. J.; Wales, J. H.; Sinnhuber, R. O.; Pawlowski, N. E. Aflatoxicol-induced hepatocellular carcinomas in rainbow trout (Salmo gairdneri) and the synergistic effects of cyclopropenoid fatty acids. Cancer Res. 41:1011–1014; 1981.

    PubMed  CAS  Google Scholar 

  18. Sinnhuber, R. O.; Hendricks, J. D.; Wales, J. H.; PutnamG.B. Neoplasms in rainbow trout, a sensitive animal model for environmental carcinogenesis. Ann. NY Acad. Sci. 298:389–408; 1977.

    Article  CAS  Google Scholar 

  19. Wales, J. H.; Sinnhuber, R. O.; Hendricks, J. D.; Nixon, J. E.; Eisele, J. A. Aflatoxin B1 induction of hepatocellular carcinoma in the embryos of rainbow trout (Salmo gairdneri. JNCI 60:1133–1139; 1978.

    PubMed  CAS  Google Scholar 

  20. Walton, M. J.; Cowey, C. B. Gluconeogenesis by isolated hepatocytes from rainbow troutSalmo gairdneri. Comp. Biochem. Physiol. 62B:75–79; 1979.

    CAS  Google Scholar 

  21. Whitham, M.; Nixon, J. E.; Sinnhuber, R. O. Liver DNA boundin vivo with aflatoxin B1 as a measure of hepatocarcinoma initiation in rainbow trout. JNCI 68:623–628; 1982.

    PubMed  CAS  Google Scholar 

  22. Williams, G. M.; Bermudez, E.; Scaramuzzino, D. Rat hepatocyte primary cell cultures. Improved dissociation and attachment technologies and the enhancement of survival by culture medium. In Vitro 13:809–817; 1978.

    Article  Google Scholar 

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Authors and Affiliations

  1. Department of Pathology, University of Maryland School of Medicine, 21201, Baltimore, MD

    Michael M. Lipsky, Talia R. Sheridan, Richard O. Bennett & Eric B. May

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  1. Michael M. Lipsky
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  2. Talia R. Sheridan
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  3. Richard O. Bennett
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  4. Eric B. May
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Additional information

This work was supported in part by research grant R809599010 from the U. S. Environmental Protection Agency.

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Lipsky, M.M., Sheridan, T.R., Bennett, R.O. et al. Comparison of trout hepatocyte culture on different substrates. In Vitro Cell Dev Biol 22, 360–362 (1986). https://doi.org/10.1007/BF02623411

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  • DOI: https://doi.org/10.1007/BF02623411

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