Skip to main content

Rat liver epithelial cell cultures in a serum-free medium: Primary cultures and derived cell lines expressing differentiated functions

In Vitro volume 20, pages 780–795 (1984)Cite this article

Summary

Rat liver epithelial cells explanted in a serum-free medium (SFM) composed of Ham's F10 basal medium plus free fatty acids adsorbed on bovine albumin gave successful rise to primary cultures and then to long-term cell lines that expressed liver functions; induction ofl-tyrosine aminotransferase by glucocorticoids, hepatic pattern of progesterone metabolism, and biosynthesis of murine primary bile acids; chenodeoxycholic and cholic acid common to higher vertebrates and α-muricholic acid specific of the rat bile.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

43,34 €

Price includes VAT (Finland)
Tax calculation will be finalised during checkout.

References

  1. Armato, U.; Draghi, E.; Andreis, P. G.; Meneghelli, V. Stimulation by N6,O2-dibutyryladenosine 3′,5′-cyclicmonophosphate of RNA and DNA synthesis and of cell proliferation of rat hepatocytes in primary tissue culture. J. Cell. Physiol. 89: 157–170; 1976.

    PubMed  Article  CAS  Google Scholar 

  2. Batta, A. K.; Salen, G.; Shefer, S. Thin-layer chromatography of conjugated bile acids. J. Chromatogr. 168: 557–561; 1979.

    PubMed  Article  CAS  Google Scholar 

  3. Bausher, J.; Schaeffer, W. I. A diploid rat liver cell culture. I. Characterization and sensitivity to aflatoxin B1. In Vitro 9: 286–293; 1974.

    Article  CAS  Google Scholar 

  4. Bègue, R. J.; Morinière, M.; Padieu, P. Urinary excretion of 5β-pregnan-3α,6α,20-triol in human gestation. J. Steroid Biochem. 9: 779–784; 1978.

    PubMed  Article  Google Scholar 

  5. Bissel, D. M.; Hammaker, L. E.; Meyer, U. A. Parenchymal cells from adult rat liver in nonproliferative monolayer culture. J. Cell Biol. 59: 722–734; 1974.

    Article  Google Scholar 

  6. Björkhem, J.; Eriksson, H.; Gustafsson, J. A.; Karlman, K. E.; Stenberg, A. Steroid hormone metabolism in developing rats. Eur. J. Biochem. 27: 318–326; 1972.

    PubMed  Article  Google Scholar 

  7. Björkhem, J.; Danielsson, H.; Wikvall, K. Hydroxylations of bile acids by reconstituted system from rat liver microsomes. J. Biol. Chem. 249: 6439–6445; 1975.

    Google Scholar 

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

    Article  CAS  Google Scholar 

  9. Bournot, P.; Maume, B. F.; Padieu, P. Sexlinked specificity of the metabolism of steroids in rats. Mass fragmentography as a method to the assay of hydrogenated metabolites of corticosterone in the liver. Biomed. Mass Spectrom. 1: 29–39; 1974.

    PubMed  Article  CAS  Google Scholar 

  10. Bournot, P.; Chessebeuf, M.; Maume, G.; Olsson, A.; Maume, B. F.; Padieu, P. Application of mass fragmentography to the study of sexlinked metabolism of testosterone and corticosterone in liver organ and in liver cells in culture. Frigerio, A.; Castagnoli, N., eds. Mass spectrometry in biochemistry and medicine. New York: Raven Press; 1974: 151–163.

    Google Scholar 

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

    PubMed  Article  CAS  Google Scholar 

  12. Chessebeuf, M.; Exilie, M. F.; Maume, G.; Piard, G.; Padieu, P. Enzymes of liver tissue. Study with epithelial liver cell culture. Analysis by cytogenetic and by mass spectrometric methods. Pharmacological applications. C. R. Soc. Biol. (Paris) 173: 469–482; 1979.

    CAS  Google Scholar 

  13. Chessebeuf, M.; Exilie, M. F.; Padieu, P.; Maume, B. F.; Morizot, J. P.; Wülfert, E. Combination of quadrupolar GC-MS techniques and cell culture systems from rodent and human liver for drug metabolism and pharmacology. In: Frigerio, A., ed. Recent developments in mass spectrometry in biochemistry and medicine and environmental research. vol 7. Amsterdam: Elsevier Sci. Pub. Co.; 1981: 123–145.

    Google Scholar 

  14. Chessebeuf, M.; Padieu, P. Institut National de la Santé et de la Recherche Médicale. Culture de cellules épithéliales en milieu sans sérum. French patent deposit no D83-04843; March 24, 1983 and P.C.T. deposit (Europe, USA and Japan) no 84/00080; March 23, 1984.

  15. Chessebeuf, M.; Fischbach, M.; Padieu, P. Time course study ofl-tyrosine aminotransferase induction in rat liver cell lines growing in serum-supplemented and serum-free media. Cell Biol. Toxicol. 1: 19–24; 1984.

    Article  Google Scholar 

  16. Davis, R. A.; Hyde, P. M.; Kuan, J. C. W.; Malone-McNeal, M.; Archambault-Schexnayder, J. Bile acid secretion by cultured rat hepatocytes. Regulation by cholesterol availability. J. Biol. Chem. 258: 3661–3667; 1983.

    PubMed  CAS  Google Scholar 

  17. Denef, C.; De Moor, P. Sexual differentiation of steroid metabolizing enzymes in the rat liver. Further studies on predetermination at birth. Endocrinology 91: 374–384; 1972.

    PubMed  CAS  Article  Google Scholar 

  18. Desgrès, J.; Guiguet, M.; Bègue, R. J.; Padieu, P. Study of progesterone metabolism in fetal and postnatal rat liver cells in culture. In: Frigerio, A.; Castagnoli, N., eds. vol 1. New York: Spectrum Publishers Inc.; 1976: 139–155.

    Google Scholar 

  19. Desgrès, J.; Fay, L.; Jo, D. H.; Guiguet, M.; Padieu, P. Oxidoreductive and hydroxylating metabolism of progesterone in rat liver epithelial cell lines. I. Metabolic pathways in a chemically defined incubation medium. J. Steroid Biochem. In press.

  20. Diamondstone, T. I. Assay of tyrosine-transaminase activity by conversion ofp-hydroxyphe-nylpyruvate top-hydroxyphenylbenzaldehyde. Anal. Biochem. 16: 395–401; 1966.

    Article  CAS  Google Scholar 

  21. Eriksson, H.; Gustafsson, J. A.; Poussette, A. Metabolism of androstenedione and progesterone in the isolated perfused rat liver. Eur. J. Biochem. 27: 327–334; 1972.

    PubMed  Article  CAS  Google Scholar 

  22. Fotherby, K. Metabolism of progesterone to 6-oxygenated steroidsin vivo andin vitro. Adv. Biosci. 3: 43–50; 1968.

    CAS  Google Scholar 

  23. Gerschenson, L. E.; Davidson, N. B.; Andersson, M. Hormonal regulation of rat liver cells cultured in chemically defined medium. Dexamethasone and insulin effects on different forms of tyrosine-α-oxoglutarate transaminase. Eur. J. Biochem. 41: 139–148; 1974.

    PubMed  Article  CAS  Google Scholar 

  24. Granner, D. K.; Lee, A.; Thompson, E. B. Interaction of glucocorticoid hormones and cyclic nucleotides in induction of tyrosine aminotransferase in cultured hepatoma cells. J. Biol. Chem. 252: 3891–3897; 1977.

    PubMed  CAS  Google Scholar 

  25. Grisham, J. W. Cell types in long-term propagable cultures of rat liver. In: Borek, C.; Williams, G. M., eds. Differentiation and carcinogenesis in liver cell cultures. New York: New York Acad. Sci. 1980: 128–137.

    Google Scholar 

  26. Guiguet, M.; Mack, G.; Exilie, M. F.; Truchot, R.; Padieu, P. Production de la fraction C3 du complément par des cellules épithéliales de foie de rat en culture. Effet stimulant de surnageants de lymphomonocytes circulants humains. C. R. Acad. Sci. (Paris). 297: 547–552; 1983.

    CAS  Google Scholar 

  27. Ham, R. G. An imporved nutrient solution for diploid Chinese hamster and human cell lines. Exp. Cell Res. 29: 515–526; 1963.

    PubMed  Article  CAS  Google Scholar 

  28. Hsia, S. L. Hyocholic acid and muricholic acids. Nair, P. P.; Kritchevsky, D., eds. The bile acids, chemistry, physiology and metabolism. Vol 1: Chemistry. New York: Plenum Press; 1971: 95–120.

    Google Scholar 

  29. Idoine, J. B.; Elliott, J. M.; Wilson, M. J.; Weisburger, E. K. Rat liver cells in culture: effect of storage, long-term culture and transformation on some enzyme levels. In Vitro 12: 541–553; 1976.

    PubMed  CAS  Google Scholar 

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

    PubMed  Article  CAS  Google Scholar 

  31. Iype, P. I.; Issaq, H. J.; Shaikh, B. Synthesis of primary bile acids by rat liver epithelial cell lines. J. Steroid Biochem. 16: 333–337; 1982.

    PubMed  Article  CAS  Google Scholar 

  32. Katsuta, H.; Takaoka, T. Carcinogenesis in tissue culture. I. Cultivation of normal rat liver cells. Jpn. J. Exp. Med. 33: 265–275; 1963.

    PubMed  CAS  Google Scholar 

  33. Kenney, F. T.; Flora, R. M. Induction of tyrosine-α-ketoglutarate transaminase in rat liver. I. Hormonal nature. J. Biol. Chem. 236: 2699–2702; 1961.

    PubMed  CAS  Google Scholar 

  34. Kenney, F. T. Induction of tyrosine-α-ketoglutarate transaminase in rat liver. III. Immunochemical analysis. J. Biol. Chem. 237: 1610–1614; 1962.

    PubMed  CAS  Google Scholar 

  35. Kletzien, R. F.; Pariza, M. W.; Becker, J. E.; Potter, V. R.; Butcher, F. R. Induction of aminoacid transport in primary cultures of adult rat liver parenchymal cells by insulin. J. Biol. Chem. 251: 3014–3020; 1976.

    PubMed  CAS  Google Scholar 

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

    PubMed  CAS  Google Scholar 

  37. Lambiotte, M.; Sjövall, J. Hydroxylation and sulfatation of bile acids in rat hepatoma cultures under the influence of a glucocorticoid. Biochem. Biophys. Res. Commun. 86: 1089–1095; 1979.

    PubMed  Article  CAS  Google Scholar 

  38. Lambiotte, M.; Thierry, N. Hydroxylation, sulfatation and conjugation of bile acids in rat hepatoma and hepatocyte culture under the influence of glucocorticoids. J. Biol. Chem. 255: 11324–11331; 1980.

    PubMed  CAS  Google Scholar 

  39. Leffert, H. L.; Paul, D. Studies on primary cultures of differentiated fetal liver cells. J. Cell Biol. 52: 559–568; 1972.

    PubMed  Article  CAS  Google Scholar 

  40. Lin, E. C. C.; Knox, W. E. Adaptation of the rat liver tyrosine α-keto-glutarate transaminase. Biochim. Biophys. Acta 26: 85–88; 1957.

    PubMed  Article  CAS  Google Scholar 

  41. Lin, R. C.; Snodgrass, P. J. Primary culture of normal adult rat liver cells which maintain stable urea cycle enzymes. Biochem. Biophys. Res. Commun. 64: 725–734; 1975.

    PubMed  Article  CAS  Google Scholar 

  42. Lowry, D. H.; Rosebrough, N. J.; Farr, A. L.; Randall, R. J. Protein measurement with the folin phenol reagent. J. Biol. Chem. 193: 265–275; 1951.

    PubMed  CAS  Google Scholar 

  43. Makita, M.; Wells, W. W. Quantitative analysis of fecal bile acids by gas-liquid chromatography. Anal. Biochem. 5: 523–530; 1963.

    PubMed  Article  CAS  Google Scholar 

  44. Malan-Shibley, L.; Iype, P. T. The influence of culture conditions on cell morphology and tyrosine aminotransferase levels in rat liver epithelial cell lines. Exp. Cell Res. 131: 363–371; 1981.

    PubMed  Article  CAS  Google Scholar 

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

    PubMed  Article  CAS  Google Scholar 

  46. Padieu, P.; Chessebeuf, M.; Olsson, A.; Lallemant, C. Conservation of differentiated characteristics in eukaryotes in single cell cultures. 7th Int. FEBS Meet. Abstr. 862: 1971; 298.

    Google Scholar 

  47. Padieu, P.; Hussein, N.; Maume, G.; Chessebeuf, M.; Tsaconas, C.; Morizot, J. P. Gas chromatography-mass spectrometry of bile sterols and acids: application to small sample of liver cell culture. International Conference on Chromatography and Mass Spectrometry in Biomedical Sciences, Bordighera, Italy, Abstr.; 1982: 53.

  48. Padieu, P.; Maume, G.; Hussein, N.; Chessebeuf, M.; Tsaconas, C. Biosynthesis of sterols and bile acids in rat liver cell lines. Biochim. Biophys. Acta (Accepted for publication).

  49. Pitot, H. C.; Peraino, C.; Morse, P. A., Jr.; Potter, V. R. Hepatomas in tissue culture compared with adapting liver in vivo. Natl. Cancer Inst. Monogr. 13: 229–245; 1964.

    PubMed  CAS  Google Scholar 

  50. Potter, V.R.; Watanabe, M.; Becker, J.; Pitot, H. C. Hormonal effects on enzyme activities in tissue culture and in whole animals. Adv. Enzyme Regul. 5: 303–316; 1967.

    PubMed  Article  CAS  Google Scholar 

  51. Ramirez, L. C.; Millot, C.; Maume, B. F. Sample purification using a C18-bonded reversed phase cartridge for the quantitative analysis of corticosteroids in adrenal cell culture by high performance liquid chromatography or gas-chromatography-mass spectrometry. J. Chromatogr. 229: 267–282; 1982.

    PubMed  Article  CAS  Google Scholar 

  52. Savage, C. R.; Bonney, R. J. Extended expression of differentiated functions in primary cultures of adult rat liver parenchymal cells maintained on nitrocellulose filters. I. Induction of phosphoenolpyruvate carboxykinase and tyrosine aminotransferase. Exp. Cell Res. 114: 307–315; 1978.

    PubMed  Article  CAS  Google Scholar 

  53. Shackleton, C. H. L.; Honour, J. W.; Taylor, N. F. Metabolism of fetal and neonatal adrenal steroids. J. Steroid Biochem. 11: 523–529; 1979.

    PubMed  Article  CAS  Google Scholar 

  54. Shefer, S.; Hauser, S.; Bekersky, I.; Mosbach, E. H. Feedback regulation of bile acid biosynthesis in the rat. J. Lipid Res. 10: 646–655; 1969.

    PubMed  CAS  Google Scholar 

  55. Sirica, A. E.; Richards, W.; Tsukada, Y.; Sattler, C. A.; Pitot, H. C. Fetal phenotypic expression by adult rat hepatocytes on collagen gel nylon meshes. Proc. Natl. Acad. Sci. USA 76: 283–287; 1979.

    PubMed  Article  CAS  Google Scholar 

  56. Sjövall, J.; Eneroth, P.; Ryhage, R. Mass spectra of bile acids. In: Nair, P. P.; Kritchevsky, D., eds. The bile acids. Chemistry, physiology and metabolism. vol 1: Chemistry. New York: Plenum Press; 1971; 209–248.

    Google Scholar 

  57. Tanaka, K.; Sato, M.; Tomita, Y.; Ichihara, A. Biochemical studies on liver functions in primary cultured hepatocytes of adult rats. I. Hormonal effects in cell viability and protein synthesis. J. Biochem. 84: 937–946; 1978.

    PubMed  CAS  Google Scholar 

  58. Thompson, E. B.; Tomkins, G. M.; Curran, J. F. Induction of tyrosine-α-ketoglutarate transaminase by steroid hormones in a newly established tissue culture cell line. Proc. Natl. Acad. Sci. USA 56: 296–303; 1966.

    PubMed  Article  CAS  Google Scholar 

  59. Tomkins, G. M.; Thompson, E. B.; Hayashi, S.; Gelehrter, T.; Granner, D. K.; Peterkofsky, B. Tyrosine transaminase induction in mammalian cells in tissue culture. Cold Spring Harbor Symp. Quant. Biol. 31: 349–360; 1966.

    PubMed  CAS  Google Scholar 

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

    PubMed  Article  CAS  Google Scholar 

  61. Williams, G. M.; Stromberg, K.; Kroes, R. Cytochemical and ultrastructural alterations associated with confluent growth in cell cultures of epithelial-like cells from rat liver. Lab. Invest. 29: 293–303; 1973.

    PubMed  CAS  Google Scholar 

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

    PubMed  Article  CAS  Google Scholar 

  63. Yousef, I. M.; Ho, J.; Jeejeebhoy, K. N. Bile acid synthesis in isolated rat hepatocytes. Can. J. Biochem. 56: 780–783; 1978.

    PubMed  CAS  Article  Google Scholar 

Download references

Author information

Affiliations

  1. Laboratoire de Biochimie Médicale, Unité INSERM U 208 and Centre National de Référence et d'Essai de Spectrométrie de Masse, Faculté de Médecine, 21033, DIJON Cedex, France

    Martine Chessebeuf & Prudent Padieu

Authors
  1. Martine Chessebeuf
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Prudent Padieu
    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

Chessebeuf, M., Padieu, P. Rat liver epithelial cell cultures in a serum-free medium: Primary cultures and derived cell lines expressing differentiated functions. In Vitro 20, 780–795 (1984). https://doi.org/10.1007/BF02618294

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02618294

Key words

  • rat liver epithelial cell culture
  • serum-free medium
  • serum-free cell line
  • l-tyrosine aminotransferase induction
  • bile acid biosynthesis
  • progesterone metabolism