A comparison of the CHO/HGPRT+ and the L5178Y/TK+/- mutation assays using suspension treatment and soft agar cloning: results for 10 chemicals.
The mouse lymphoma assay (MLA) and Chinese hamster ovary (CHO) cell assay are sensitive indicators of mutagenicity. The CHO assay has been modified technically to permit treatment in suspension and soft agar cloning comparable to the MLA. This methodology eliminates the risk of metabolic cooperation and the trauma of trypsinization. In addition, a larger population of cells can be treated and cloned for mutant selection. In order to compare the effectiveness of the test systems, 10 chemicals were evaluated for the induction of forward mutations in the CHO and MLA. Several of these chemicals have been reported as clastogenic; therefore, abbreviated colony sizing was performed to gauge the extent of genetic damage to the MLA cells. Both test systems detected benzo[a]pyrene, mitomycin C, acridine orange, and proflavin, and, with the exception of proflavin, more large colonies were present than small colonies. The suspect clastogen, phenytoin, was not mutagenic in the MLA and produced inconclusive results in the CHO. Ethidium bromide, a clastogen and a bacterial mutagen, was not mutagenic in either the MLA or CHO. Four compounds (p-aminophenol, benzoin, methoxychlor, and pyrene) were positive in the MLA, generally inducing a large number of small colonies, while demonstrating no mutagenic activity in the CHO assay. They have also been shown to be generally nongenotoxic in other test systems. Overall, the modified CHO assay did not appear to be better than the MLA for the detection of mutagenic agents. However, the MLA does appear to have lower specificity.
Oberly
T J
TJ
Toxicology Research Laboratories, Lilly Research Laboratories, Greenfield, IN 46140.
Michaelis
K C
KC
Rexroat
M A
MA
Bewsey
B J
BJ
Garriott
M L
ML
D000362:Agar; D000818:Animals; D016466:CHO Cells / Q000187:drug effects / Q000201:enzymology; D006224:Cricetinae; D005069:Evaluation Studies as Topic; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics*; D007940:Leukemia L5178 / Q000201:enzymology / Q000235:genetics; D051379:Mice; D009152:Mutagenicity Tests / Q000379:methods*; D009153:Mutagens / Q000494:pharmacology*; D013937:Thymidine Kinase / Q000235:genetics*; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000201:enzymology
D009153:Mutagens; D000362:Agar; D007041:Hypoxanthine Phosphoribosyltransferase; D013937:Thymidine Kinase