Skip to main content
SpringerLink
Log in

Conditions affecting primary cell cultures of functional adult rat hepatocytes

II. Dexamethasone enhanced longevity and maintenance of morphology

  • Published:
In Vitro - Plant Aims and scope Submit manuscript

Summary

Primary monolayer cell cultures of adult rat hepatocytes underwent change in morphology and substantial cell loss between 1 and 3 days postinoculation. Dexamethasone-supplementation (1µM) of the culture medium maintained the polygonal epithelial morphology of the hepatocytes and increased longevity such that over 80% of the cells survived for 3 days and at least 30% for 8 or 9 days. This enhancement of survival was obtained up to 48 hr postinoculation, but the earlier the time of dexamethasone supplementation the greater the effect. Removal of dexamethasone resulted in a decrease in longevity. The positive effect of dexamethasone on longevity was observed following dexamethasone replacement of insulin in supplemented cultures, but the combination of insulin and dexamethasone resulted in poorer survival than with dexamethasone alone. The results are interpreted to indicate that dexamethasone provided a requirement of the in vitro environment for survival and suggest that elaboration of a complex medium is required to maintain hepatocytes in culture.

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. Sandstrom, B. 1964. A method of cultivating morphologically highly differentiated liver epithelium in vitro. Acta Soc. Med. Upsal. 69:209–214.

    Google Scholar 

  2. Rose, G. G., M. Kumegawa, and M. Cattoni. 1968. The circumfusion system for multipurpose culture chambers. II. The protracted maintenance of differentiation of fetal and newborn mouse liver in vitro. J. Cell Biol. 39:430.

    Article  PubMed  CAS  Google Scholar 

  3. Leffert, H. L., and D. Paul. 1972. Studies of primary cultures of differentiated fetal liver cell. J. Cell Biol. 52:559–568.

    Article  PubMed  CAS  Google Scholar 

  4. Parsa, I., and L. Flancbaum. 1975. Long term organ culture of rat liver rudiment in a synthetic medium: morphological and biochemical development. Dev. Biol. 4:120–131.

    Article  Google Scholar 

  5. Michalopoulos, G., and H. C. Pitot. 1975. Primary culture of parenchymal liver cells on collagen membranes. Morphological and biochemical observations. Exp. Cell Res. 94:70–78.

    Article  PubMed  CAS  Google Scholar 

  6. Katsuta, H., and T. Takaoka. 1963. Carcinogenesis in tissue culture. II. Cultivation of normal rat liver cells. Japan J. Exp. Med. 33:265–275.

    CAS  Google Scholar 

  7. Biberfeld, P., T. L. E. Ericsson, P. Perlman, and M. Z. Raftell. 1966. Ultrastructural features of in vitro propagated rat liver cells. Z. Zellforsch. 71:153–168.

    Article  PubMed  CAS  Google Scholar 

  8. Gerschenson, L. E., and D. Casanello. 1968. Metabolism of rat liver cells cultured in suspension: insulin and glucagon effects on glycogen level. Biochem. Biophys. Res. Comm. 33:584–589.

    Article  PubMed  CAS  Google Scholar 

  9. Coon, H. 1969. Clonal culture of differentiated cells from mammals: rat liver cell culture. Carnegie Inst. Washington Year Book 67:419–421.

    Google Scholar 

  10. Borek, C., S. Higashino, and W. R. Lowenstein. 1969. Intercellular communication and tissue growth. IV. Conduction of membrane junctions of normal and cancerous cells in culture. J. Membr. Biol. 1:274–293.

    Article  Google Scholar 

  11. Waymouth, C., H. W. Chen, and B. G. Wood. 1971. Characteristics of mouse liver parenchymal cells in chemically defined media. In Vitro 6:371.

    Article  Google Scholar 

  12. Kaighn, M. E., and A. M. Prince. 1971. Production of albumin and other serum proteins by clonal cultures of normal human liver. Proc. Natl. Acad. Sci. U.S.A. 68:2396–2400.

    Article  PubMed  CAS  Google Scholar 

  13. Iype, P. T. 1971. Cultures from adult rat liver cells. I. Establishment of monolayer cell-culture from normal liver. J. Cell. Physiol. 78:281–288.

    Article  PubMed  CAS  Google Scholar 

  14. Rutzky, L. P., and W. G. Taylor, and R. W. Pumper. 1971. In vitro stimulation of rat liver cells by homologous partial hepatectomy serum. In Vitro 7:1–5.

    Article  PubMed  CAS  Google Scholar 

  15. Williams, G. M., E. K. Weisburger, and J. H. Weisburger. 1971. Isolation and long-term cell culture of epithelial-like cells from rat liver. Exp. Cell Res. 69:106–112.

    Article  PubMed  CAS  Google Scholar 

  16. Diamond, L., R. McFall, Y. Tashiro, and D. Sabatini. 1973. The WIRL-3 rat liver cell lines and their transformed derivatives. Cancer Res. 33:2627–2636.

    PubMed  CAS  Google Scholar 

  17. Chessebeuf, M., A. Olsson, P. Bournot, J. Desgres, M. Guiguet, G. Maume, B. F. Maume, B. Perissel, and P. Padieu. 1974. Long-term cell culture of rat liver epithelial cells retaining some hepatic functions. Biochimie 56:1365–1379.

    Article  PubMed  CAS  Google Scholar 

  18. Bausher, J., and W. I. Schaeffer. 1974. A diploid rat liver cell culture. I. Characterization and sensitivity to aflatoxin B. In Vitro 9:286–293.

    Article  CAS  Google Scholar 

  19. Masuji, H., H. Nakabayashi, and J. Sato. 1974. Long-term cultivation of two diploid epithelial cell lines derived from normal rat liver cells. Acta Med. (Okayama) 28:281–293.

    CAS  Google Scholar 

  20. Williams, G. M., and J. M. Gunn. 1974. Long-term cell culture of adult rat liver epithelial cells. Exp. Cell Res. 89:139–142.

    Article  PubMed  CAS  Google Scholar 

  21. Seglen, P. O. 1973. Preparation of rat liver cells. III. Enzymatic requirements for tissue dispersion. Exp. Cell Res. 82:391–398.

    Article  PubMed  CAS  Google Scholar 

  22. Laishes, B. A., and G. M. Williams. 1976. Conditions affecting primary cell cultures of functional adult rat hepatocytes. I. The effect of insulin. In Vitro, 12: 521–532.

    PubMed  CAS  Google Scholar 

  23. Williams, G. M. 1976. Primary and long-term culture of adult rat liver epithelial cells. In: D. M. Prescott (Ed.),Methods in Cell Biology. Volume 14: 357–364.

    Article  PubMed  CAS  Google Scholar 

  24. Bonney, R. J., J. E. Becker, P. R. Walker, and V. R. Potter. 1974. Primary monolayer cultures of adult rat liver parenchymal cells suitable for study of the regulation of enzyme synthesis. In Vitro 9:399–413.

    Article  CAS  Google Scholar 

  25. Gunn, J. M., H. Shinozuka, and G. M. Williams. 1976. Enhancement of phenotypic expression in cultured malignant liver epithelial cells by a complex medium. J. Cell. Physiol. 87:79–88.

    Article  PubMed  CAS  Google Scholar 

  26. Arpels, C., V. I. Babcock, and C. M. Southam. 1964. Effect of steroids on human cell cultures; sustaining effects of hydrocortisone. Proc. Soc. Exp. Biol. Med. 115:102–106.

    PubMed  CAS  Google Scholar 

  27. Yuan, G. C., and R. S. Change, J. B. Little, and G. Cornil. 1967. Prolongation of postmitotic lifespan of primary human amnion cells in vitro by hydrocortisone. J. Gerontol. 22:174–179.

    PubMed  CAS  Google Scholar 

  28. Macieira-Coelho, A. 1966. Action of cortisone on human fibroblasts in vitro. Experientia 22:390–391.

    Article  PubMed  CAS  Google Scholar 

  29. Smith, B. T., J. S. Torday, and C. J. P. Giroud. 1973. The growth promoting effect of cortisol on human fetal lung cells. Steroids 22:515–524.

    Article  PubMed  CAS  Google Scholar 

  30. Cristofalo, V. J., and J. Kabakjian. 1975. Lysosomal enzymes and aging in vitro: subcellular enzyme distribution and effect of hydrocortisone on cell life-span. Mech. Ageing Dev. 4:19–28.

    Article  PubMed  CAS  Google Scholar 

  31. Farber, J. L., and S. K. El-Mofty. 1975. The biochemical pathology of liver cell necrosis. Am. J. Pathol. 81:237–250.

    PubMed  CAS  Google Scholar 

  32. Ballard, P. L., and G. M. Tomkins. 1969. Hormone modification of the cell surface. Nature 224:344–345.

    Article  PubMed  CAS  Google Scholar 

  33. Billat, C., C. Plas, and R. Jacquot. 1972. Etude en microscopie electronique d’hepatocytes foetaux de rat en culture primaire: action du cortisol. Compt. Rend. 274:2338–2340.

    CAS  Google Scholar 

  34. Lambiotte, M., A. Vorbrodt, and E. L. Benedetti, 1973. Expression of differentiation of rat foetal hepatocytes in cellular culture under the action of glucocorticoids: appearance of the canaliculi. Cell Diff. 2:43–53.

    Article  CAS  Google Scholar 

  35. Berliner, J. A. 1975. Effect of glucocorticoids on the ultrastructure of cultured liver cells. In: L. E. Gerschenson, and E. B. Thompson (Eds.),Gene Expression and Carcinogenesis in Cultured Liver. Academic Press, New York, pp. 62–93.

    Google Scholar 

  36. Gelehrter, T. D., W. L. Risser, and S. B. Reichberg. 1975. Hormonal regulation of amino acid transport in rat hepatoma cells in tissue culture. In: L. E. Gerschenson, and E. B. Thompson (Eds.),Gene Expression and Carcinogenesis in Cultured Liver. Academic Press, New York, pp. 190–204.

    Google Scholar 

  37. Kletzien, R. F., M. W. Pariza, J. E. Becker, and V. R. Potter. 1975. A “permissive” effect of dexamethasone on the glucagon induction of amino acid transport in cultured hepatocytes. Nature 256:46–47.

    Article  PubMed  CAS  Google Scholar 

  38. Harper, H. A. 1973.Review of Physiological Chemistry. Fourteenth edition. Lange Medical Publications, Los Altos, Calif.

    Google Scholar 

Download references

Author information

Author notes

  1. Brian A. Laishes (Research Fellow of the National Cancer Institute of Canada)

    Present address: Department of Pathology, University of Toronto, Medical Sciences Building, M5S 1A8, Toronto, Canada

Authors and Affiliations

  1. Fels Research Institute and Department of Pathology, Temple University School of Medicine, 19140, Philadelphia, Pennsylvania

    Brian A. Laishes (Research Fellow of the National Cancer Institute of Canada) & Gary M. Williams

Authors
  1. Brian A. Laishes
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Gary M. Williams
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

This study was supported by an Alexander Ralston Peacock Memorial Grant for Cancer Research (No. BC-133A) from the American Cancer Society.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Laishes, B.A., Williams, G.M. Conditions affecting primary cell cultures of functional adult rat hepatocytes. In Vitro Cell.Dev.Biol.-Plant 12, 821–832 (1976). https://doi.org/10.1007/BF02796367

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02796367

Key words

Search

Navigation