Utilization of an established rat hepatoma cell line for mutation studies

A selected strain of rat hepatoma cells was evaluated for its utility in the assay of mutation. We demonstrate that these cells have practical and theoretical advantages over most other mammalian cells utilized for the mutation assay at the HGPRT locus. Characteristics of the H4IIE cell line which enhance the ability of the HGPRT assay to detect mutation include: intrinsic metabolic capabilities; small cell size, and lack of mobility; colony-forming efficiency of 85–95%; a background mutation level between 0.9 and 3 mutants per 10⁵ viable cells; and the ability to proliferate in medium containing 5% fetal bovine serum and 5% horse serum. The optimal expression period for the maximum frequency of mutants was investigated using UVC as the mutagenic agent, and found to plateau after 8–10 days. The metabolic capacity of this cell line was demonstrated using 2-aminoanthracene and cyclophosphamide, two types of mutagens requiring biotransformation for activity. We conclude that the use of the H4/HGPRT system could prove valuable in the mutagenic screening of suspect environmental agents.