In Vitro

, Volume 9, Issue 6, pp 399–413

Primary monolayer cultures of adult rat liver parenchymal cells suitable for study of the regulation of enzyme synthesis

  • Robert J. Bonney
  • Joyce E. Becker
  • P. Roy Walker
  • R. Van Potter
Article

Summary

Parenchymal cells from adult rat liver which had been fully regenerated were isolated and cultured in nonproliferating monolayers in vitro. The optimum conditions for attachment of these cells to Falcon plastic dishes were determined. When approximately 1.0×105 nuclei per cm2 suspended in Ham's F-12 medium with 0.5 μg of insulin per ml and 25% fetal calf serum were incubated at 37°C for 24 hr, about 50% became attached and contiguous. When the above medium was supplemented with synthetic buffers 2-(N-morpholino) ethanesulfonic acid (MES) andN-tris (hydroxymethyl) methyl-2-aminoethanesulfonic acid (TES), the presence of 15% fetal calf serum also allowed an attachement effiency of 50%. Tyrosine aminotrasferase activity in the cells was elevated when the culture medium was supplemented with hydrocortisone or dexamethasone. The largest increases were observed after 72 hr of culture. Cycloheximide prevented the increase.

Key words

primary culture rat liver liver parenchymal cells enzyme regulation tyrosine aminotransferase glucocorticoids 

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Copyright information

© Tissue Culture Association 1974

Authors and Affiliations

  • Robert J. Bonney
    • 1
  • Joyce E. Becker
    • 1
  • P. Roy Walker
    • 1
  • R. Van Potter
    • 1
  1. 1.McArdle Laboratory for Cancer ResearchUniversity of WisconsinMadison
  2. 2.New York State Department of HealthAlbany
  3. 3.Deparment of Biochemistry, The UniversitySheffieldEngland

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