Method and cell line for testing mutagenicity of a chemical
AssigneeWayne State University
InventorsJ. Christopher States, Ronald N. Hines, Raymond F. Novak
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Abstract
A method of testing the mutagenicity of a chemical includes the steps of exposing test cells to the chemical in vitro, intracellularly metabolizing the chemical into a mutagenic or cytotoxic metabolite and then detecting gene/protein/cell damage in the test cells as an indication of the mutagenicity/cytotoxicity of the chemical. A cell line is provided for testing mutagenicity of the chemicals, the cell line consisting essentially of fibroblasts normally having no detectable cytochrome P450 mixed function oxidase enzyme activity. The fibroblasts are transformed with chimeric gene constructs containing cytochrome P450 coding sequences and have intracellular cytochrome P450 oxidative metaboli zing activity.
Claims15
A method of testing for mutagenicity of a chemical by exposing human fibroblast test cells normally not including any cytochrome P450 activity, the test cells being transformed to express cytochrome P450 enzymes through the human CYP1A1 gene to the chemical in vitro;
- intracellularly metabolizing the chemical into a mutagenic metabolite by oxidatively metabolizing the chemical within the fibroblasts through the intracellular cytochrome P450 mixed function oxidase enzymes expressed by said transformed fibroblasts;
- and detecting gene damage in the test cells as an indication of mutagenicity of the chemical.
A cell line for testing for mutagenicity of a chemical, said cell line consisting essentially of fibroblasts normally having no detectable cytochrome P450 mixed function oxidase enzyme activity, said fibroblasts being transformed with chimeric gene constructs containing cytochrome P450 coding sequences and having intracellular cytochrome P450 oxidative metabolizing activity.
A method of making a cell line for testing mutagenicity of a chemical, said method comprising the step of transforming XPA cells which are of a DNA repair deficient human fibroblast cell line, to express cytochrome P450 enzyme activity intracellularly.
Dependencies are inferred from the claim text.