In Vitro Cellular & Developmental Biology

, Volume 26, Issue 11, pp 1057–1062

Drug metabolizing enzymes in rat hepatocytes co-cultured with cell lines

  • M. Teresa Donato
  • M. José Gómez-Lechón
  • José V. Castell
Regular Papers

DOI: 10.1007/BF02624440

Cite this article as:
Donato, M.T., Gómez-Lechón, M.J. & Castell, J.V. In Vitro Cell Dev Biol (1990) 26: 1057. doi:10.1007/BF02624440

Summary

We have developed new co-cultures of continuous cell lines 3T3 (clone A31) and C3H/10T1/2 (colone 8) with hepatocytes as an alternative to co-cultures with nonconinuous epithelial cells. In this biological system we studied in detail the expression of the hepatic biotransformation system. After 7 d in culture, total cytochrome P-450 content and the monooxygenase activities aryl hydrocarbon hydroxylase and 7-ethoxycoumarino-deethylase still maintained about 30% of their initial value, whereas in pure cultured hepatocytes these activities were undetectable. A significant response to induction by methylcholanthrene and phenobarbital of monooxygenase activities was observed in co-cultures for 7 d. NADPH-cytochrome c reductase activity remained unchanged for at least 7 d in co-cultured hepatocytes, whereas in pure cultures this activity was reduced to about 75% of the initial value after only 24 h. Finally, the activity of the conjugating enzymes UDP-Gt and GSH-t was maintained at nearly the initial levels during the complete period of study. The easy handling of continuous cell lines and the maintenance of the biotransformation system of hepatocytes in co-culture make this approach simpler and easier to standardize.

Key words

co-cultureshepatocytescytochrome P-450aryl hydrocarbon hydroxylase7-ethoxycoumarino-deethylaseconjugating enzymes

Copyright information

© Tissue Culture Association 1990

Authors and Affiliations

  • M. Teresa Donato
    • 1
  • M. José Gómez-Lechón
    • 1
  • José V. Castell
    • 1
  1. 1.Unidad de Hepatología Experimental, Centro de InvestigaciónHospital La Fe, Serivicio Valenciano de SaludValenciaSpain