Mutagenesis, Vol. 14, No. 3, 265-270,
May 1999
© 1999 UK Environmental Mutagen Society/Oxford University Press
Discussion Forum |
The mouse lymphoma assay in the wake of ICH4—where are we now?
MRC Cell Mutation Unit, University of Sussex, Falmer, Brighton, East Sussex BN1 9RR, UK and 1 Environmental Carcinogenesis Division, National Health and Environmental Effects Research Laboratory, US Environmental Protection Agency, Research Triangle Park, NC 27711, USA
Background
The ultimate goal of short-term tests should surely be to detect the complete spectrum of heritable effects or genetic events involved in the aetiology of cancer. Thus the types of mutational events that must be detected include point mutations, deletions, translocations, rearrangements, aneuploidy and mitotic recombination/gene conversion. The mouse lymphoma assay (MLA) has been widely used for many years for determining the potential for chemicals to cause the full broad spectrum of mutational damage (Applegate et al., 1990
; Mitchell et al., 1997). Substantial research has been conducted to understand the capabilities (and limitations) of the assay, its proper conduct and the appropriate interpretation of test data (Clive et al., 1979, 1990; Hozier et al., 1981, 1982, 1989; Moore-Brown, 1981; Cole et al., 1983, 1990, 1991; Turner et al., 1984; Moore et al
Overview of MLA development and assay characterization
Phase I. Initial protocol development
Phase II. Resolution of protocol issues, evaluation of the ability of the MLA to detect chemical clastogens and aneugens and detailed mutant analysis
Protocol improvement and the use of a microwell cloning procedure. Comparison of chromosome aberration frequency with small-colony mutant induction. Chromosomal analysis of large and small mutant colonies. The NcoI restriction fragment length polymorphism (RFLP) analysis. p53 status of L5178y mouse lymphoma cells. The importance of optimal small colony mutant detection. ICH4 and the Japanese Trial: is a new protocol justified?
Agar versus microwell cloning
The requirement for 24 h treatment to detect some aneugens
Validation of the ability of the microwell/MLA to detect aneugens
Summary and recommendatons
Notes
References
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