Quantitative assay for albumin-producing liver cells after simian virus 40 transformation of rat hepatocytes maintained in chemically defined medium.

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Abstract
Transformation of rat hepatocytes by SV40 in chemically defined medium was examined. When hepatocytes plated on collagen-coated plates were infected with SV40, colonies of replicating cells appeared as early as 40 days after infection; no colonies appeared in control cultures. Medium from 85% of the transformed cultures contained albumin. When collagen was eliminated and heptocytes were plated on Primaria plastic cell culture dishes, transformation occurred; medium from 86% of the transformed cultures contained albumin but the maximum albumin level secreted per culture was only 62% of that produced by cultures on collagen-coated plants. Quantitative assays for transformation were established. Transformation was linear after infection with 2-50 plaque-forming units of virus per hepatocyte, and the transformation frequency was the same on the 2 plating surfaces. An immuno-overlay technique made it possible to identify, purify and determine the morphology of the albumin-producing cells. When ornithine was substituted for arginine in the medium, the transformation frequency decreased markedly while the percentage of colonies producing albumin increased from 30 to 100%. An assay for quantifying transformation of a normal hepatocyte population and for identifying and enumerating epithelial liver cell transformants that produce albumin was defined.