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US 4,302,535 AGranted
Assay for mutagenesis in heterozygous diploid human lymphoblasts
AssigneeMassachusetts Institute of Technology
InventorsThomas R. Skopek, Howard L. Liber, Bruce W. Penman, William G. Thilly, Henry Hoppe, IV
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Abstract
An assay is disclosed for determining mutagenic damage caused by the administration of a known or suspected mutagen to diploid human lymphoblastoid cell lines. The gene locus employed for this assay is the gene for thymidine kinase, uridine kinase, or cytidine deaminase. Since human lymphoblastoid cells contain two genes for these enzymes, heterozygotes of human lymphoblastoid cells are used in this assay.
Claims7
An assay for determining mutagenesis, comprising:
- a. exposing a culture of human diploid or near-diploid cells capable of continuous division in suspension culture, said cells being heterozygotes having only one active gene for an enzyme for converting a pyrimidine analog to a toxic material for said cells to an agent to be tested for its mutagenic effects on said cells;
- b. incubating exposed cells for a number of generations to allow full expression of phenotypically-developed resistance to a pyrimidine analog which acts as a substrate for said enzyme and is normally toxic to said cells;
- c. culturing cells having the full phenotypically developed resistance both in the presence of and without a pyrimidine analog which is toxic to non-mutant cells;
- and,
- d. comparing the amount of mutant cells to the amount of non-mutant cells to thereby determine the degree of induced mutagenesis.
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Dependencies are inferred from the claim text.