Assay for mutagenesis in diploid human lymphoblasts

G - Physics – 01 – N

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G01N 33/48 (2006.01) G01N 33/50 (2006.01)

Patent

CA 1091562

ASSAY FOR MUTAGENESIS IN DIPLOID HUMAN LYMPHOBLASTS ABSTRACT OF THE DISCLOSURE An assay is disclosed for determining mutagenic damage caused by the administration of a known or suspected mutagen to diploid human lymphoblastoid cell lines. After administration of or exposure to a mutagenic agent, the lymphoblasts are incubated for a sufficient number of genera- tions to allow full expression of phenotypic resistance to 6-thioguanine or other purines which serve as substrates for hypoxanthine guanine phosphoribosyl transferase (HGPRT). A surprising discovery is the remarkably long length of time required for such phenotypic expression. After the pheno- typic lag has passed, the mutant fraction can be determined to complete the assay. A nontoxic, active, sterile micro- somal drug-metabolizing system compatible with mammalian cell bioassays is also disclosed which can be used in the assay to determine metabolite-caused mutagenesis. These bioassay systems can be used by genetic toxicologists to determine the potential genetic hazards to human beings for a variety of suspected or known mutagens, including newly developed chemicals.

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