Primary Monolayer Cultures of Adult Rat Liver Parenchymal Cells Suitable for Study of the Regulation of Enzyme Synthesis

• Published in: In Vitro Vol. 9; no. 6; pp. 399 - 413
• Main Authors: Bonney, Robert J., Becker, Joyce E., Walker, P. Roy, Potter, Van R.
• Format: Journal Article
• Language: English
• Published: United States Tissue Culture Association, Inc 01-05-1974
• Subjects: Alkanesulfonates; Animals; Autoradiography; Blood; Buffers; Cell Adhesion; Cell culture techniques; Cell Division; Cell lines; Cell Separation; Cells, Cultured; Culture Media; Cultured cells; Cycloheximide - pharmacology; Dexamethasone - pharmacology; DNA - biosynthesis; Enzymes; Epithelioid cells; Hepatocytes; Hydrocortisone - pharmacology; Insulin; Liver; Liver - cytology; Liver - enzymology; Liver cells; Liver Regeneration; Male; Mesylates; Methanol; Microscopy, Phase-Contrast; Morpholines; Rats; Serum Albumin, Bovine; Thymidine - metabolism; Tritium; Tyrosine Transaminase - biosynthesis
• ISSN: 0073-5655; 2327-4328; 1475-2689
• DOI: 10.1007/bf02615992

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$1.0 \times 10^5 nuclei per cm^2$suspended in Ham's F-12 medium with$0.5 \mu g$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) and N-tris (hydroxymethyl) methyl-2-aminoethanesulfonic acid (TES), the presence of 15% fetal calf serum also allowed an attachment efficiency of 50%. Tyrosine aminotransferase 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.