ASTM E1280-97(2008)
(Guide)Standard Guide for Performing the Mouse Lymphoma Assay for Mammalian Cell Mutagenicity (Withdrawn 2015)
Standard Guide for Performing the Mouse Lymphoma Assay for Mammalian Cell Mutagenicity (Withdrawn 2015)
SIGNIFICANCE AND USE
This guide is limited to procedures used solely for the testing of substances to determine their mutagenicity and does not apply to other methods and uses such as exploring mechanisms of mutation.
Recent evidence suggests that this assay measures a dual genetic end point; therefore, some discussion of the relationships between mammalian cell mutagenicity testing results and the results observed both in pure gene mutational assays and in cytogenetic assays is necessary. However, it is not the intent of this guide to discuss other relationships between this mammalian cell mutagenicity testing results and the results observed in other tests for mutagenicity and carcinogenicity.
SCOPE
1.1 The purpose and scope of this guide is to present background material and to establish criteria by which protocols and procedures for conducting the L5178Y/TK+/−-3.7.2C mouse lymphoma mutagenicity assay (commonly referred to as the mouse lymphoma assay, (MLA)) can be properly understood and evaluated. This guide is also intended to aid researchers and others to gain a better understanding of the critical elements involved with mammalian cell mutagenicity testing. More specifically, this guide is intended to provide for researchers the accomplishment of the following goals:
1.1.1 Provide an understanding of the critical procedures (steps) in the performance of this mammalian cell mutagenicity test.
1.1.2 Provide generalized criteria by which researchers can evaluate if they are properly performing, utilizing, and interpreting this assay.
1.1.3 Provide criteria by which individuals responsible for evaluating MLA data can determine if the experiments have been properly performed and interpreted.
1.1.4 Provide a basis from which new procedures and developments in testing procedures can be evaluated.
1.1.5 Provide an understanding of the types of genetic damage (that is, gene and chromosome mutation) that may be detected in this mammalian cell mutagenicity test.
1.2 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.
WITHDRAWN RATIONALE
The purpose and scope of this guide was to present background material and to establish criteria by which protocols and procedures for conducting the L5178Y/TK+/−-3.7.2C mouse lymphoma mutagenicity assay (commonly referred to as the mouse lymphoma assay, (MLA)) could be properly understood and evaluated.
Formerly under the jurisdiction of Committee F04 on Medical and Surgical Materials and Devices, this guide was withdrawn in November 2014. This standard is being withdrawn without replacement due to its limited use by industry.
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NOTICE: This standard has either been superseded and replaced by a new version or withdrawn.
Contact ASTM International (www.astm.org) for the latest information
Designation: E1280 − 97(Reapproved 2008)
Standard Guide for
Performing the Mouse Lymphoma Assay for Mammalian
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Cell Mutagenicity
This standard is issued under the fixed designation E1280; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision.Anumber in parentheses indicates the year of last reapproval.A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
INTRODUCTION
This guide was developed at the request of ASTM Subcommittee E47.09 on Biomarkers in order
to aid toxicologists, geneticists, biochemists, other researchers, and interested persons in the
understanding, performance, and analysis of the mammalian cell mutagenicity test that uses the
+/−
TK -3.7.2C strain of L5178Ymouse lymphoma cells. In this rapidly changing area of toxicology, it
is not intended for this guide to replace, alter, or diminish the usefulness of presently available
protocols and procedures.
1. Scope 1.2 The values stated in SI units are to be regarded as
standard. No other units of measurement are included in this
1.1 The purpose and scope of this guide is to present
standard.
background material and to establish criteria by which proto-
+/−
cols and procedures for conducting the L5178Y/TK -3.7.2C
2. Terminology
mouselymphomamutagenicityassay(commonlyreferredtoas
2.1 Definitions:
the mouse lymphoma assay, (MLA)) can be properly under-
2.1.1 clastogen—any agent that is capable of inducing
stood and evaluated. This guide is also intended to aid
chromosome breaks.
researchers and others to gain a better understanding of the
2.1.2 gene mutation—any heritable change whose physical
critical elements involved with mammalian cell mutagenicity
extent is restricted to the limits of a single gene.
testing. More specifically, this guide is intended to provide for
researchers the accomplishment of the following goals:
2.1.3 mutagen—any physical or chemical agent capable of
1.1.1 Provide an understanding of the critical procedures inducing a mutation.
(steps)intheperformanceofthismammaliancellmutagenicity
2.1.4 mutation—any heritable change in the genetic
test.
material, not caused by genetic segregation or genetic
1.1.2 Provide generalized criteria by which researchers can recombination, and that is transmitted to daughter cells.
evaluate if they are properly performing, utilizing, and inter-
2.2 Definitions of Terms Specific to This Standard:
preting this assay.
2.2.1 chromosome mutation—a mutation resulting from a
1.1.3 Provide criteria by which individuals responsible for
structural change to a chromosome involving the gain, loss, or
evaluating MLA data can determine if the experiments have
relocation of chromosome segments. Chromosome mutations
been properly performed and interpreted.
can be either intrachromosomal or interchromosomal.
1.1.4 Provide a basis from which new procedures and
2.2.2 relative suspension growth (RSG)—used to measure
developments in testing procedures can be evaluated.
the cytotoxicity of a given treatment based on the growth of
1.1.5 Provide an understanding of the types of genetic
cells in suspension culture relative to the untreated or solvent
damage (that is, gene and chromosome mutation) that may be control(s). RSG is calculated according to the method of Clive
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detected in this mammalian cell mutagenicity test.
and Spector (1).
2.2.3 relative total growth (RTG)—used as a means to
measure the relative toxicity to cells (survival) following
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treatment in the mouse lymphoma assay. RTG is calculated
This guide is under the jurisdiction of Committee F04on Medical and Surgical
Materials and Devices and is the direct responsibility of Subcommittee F04.16 on
Biocompatibility Test Methods.
Current edition approved Aug. 1, 2008. Published August 2008. Originally
2
approved in 1989. Last previous edition approved in 2003 as E1280–97 (2003). Theboldfacenumbersinparenthesesrefertothelistofreferencesattheendof
DOI: 10.1520/E1280-97R08. this guide.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
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E1280 − 97 (2008)
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according to the method of Clive and Spector (1) and includes usedforavarietyofmediapreparations.PluronicF68 mustbe
RSG as well as the ability to form colonies in the clonal phase added to the base medium to facilitate growth in suspension
culture. Other supplements usually include antibiotics, sodium
of the assay.
pyruvate, and occasionally, glutamine. Refer to references in
2.3 Symbols:
4.1 for suggested concentrations.
2.3.1 BrUdR—5-bromo-2`-deoxyuridine.
4.1.2 Growth Medium—Growth medium is prepared by
supplementing the base medium with horse serum, usually
2.3.2 BUdR—bromouracil deoxyriboside.
10%
...
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