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

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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.

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Author notes

  1. Robert J. Bonney

    Present address: New York State Department of Health, New Scotland Ave., 12201, Albany, N. Y.

  2. P. Roy Walker

    Present address: Deparment of Biochemistry, The University, S10 2TN, Sheffield, England

Authors and Affiliations

  1. McArdle Laboratory for Cancer Research, University of Wisconsin, 53706, Madison, Wisconsin

    Robert J. Bonney, Joyce E. Becker, P. Roy Walker & R. Van Potter

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  1. Robert J. Bonney
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  2. Joyce E. Becker
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  3. P. Roy Walker
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  4. R. Van Potter
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Additional information

The work was supported in part by National Cancer Institute Grants CA-51304-01 (R. J. B.) and CA-07175. P. R. W. was a Damon Runyon Memorial Fund Postdoctoral Fellow.

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Bonney, R.J., Becker, J.E., Walker, P.R. et al. Primary monolayer cultures of adult rat liver parenchymal cells suitable for study of the regulation of enzyme synthesis. In Vitro 9, 399–413 (1974). https://doi.org/10.1007/BF02615992

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