ASTM E1280-97(2003)

Standard Guide for Performing the Mouse Lymphoma Assay for Mammalian Cell Mutagenicity

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.

General Information

• Status: Historical
• Publication Date: 09-Sep-2003
• ICS: 07.100.10 - Medical microbiology
• Technical Committee: F04 - Medical and Surgical Materials and Devices
• Drafting Committee: F04.16 - Biocompatibility Test Methods

Relations

• Replaces: ASTM E1280-97 - Standard Guide for Performing the Mouse Lymphoma Assay for Mammalian Cell Mutagenicity
• Effective Date: 10-Sep-2003
• Replaced By: ASTM E1280-97(2008) - Standard Guide for Performing the Mouse Lymphoma Assay for Mammalian Cell Mutagenicity (Withdrawn 2015)
• Effective Date: 01-Aug-2008
• Referred By: ASTM F1439-03(2018) - Standard Guide for Performance of Lifetime Bioassay for the Tumorigenic Potential of Implant Materials
• Effective Date: 10-Sep-2003

Standards Content (Sample)

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 2003)
Standard Guide for
Performing the Mouse Lymphoma Assay for Mammalian
1
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 (e) 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 2.1.1 clastogen—any agent that is capable of inducing
chromosome breaks.
1.1 The purpose and scope of this guide is to present
2.1.2 gene mutation—any heritable change whose physical
background material and to establish criteria by which proto-
+/−
extent is restricted to the limits of a single gene.
cols and procedures for conducting the L5178Y/TK -3.7.2C
2.1.3 mutagen—any physical or chemical agent capable of
mouselymphomamutagenicityassay(commonlyreferredtoas
inducing a mutation.
the mouse lymphoma assay, (MLA)) can be properly under-
2.1.4 mutation—any heritable change in the genetic mate-
stood and evaluated. This guide is also intended to aid
rial, not caused by genetic segregation or genetic recombina-
researchers and others to gain a better understanding of the
tion, and that is transmitted to daughter cells.
critical elements involved with mammalian cell mutagenicity
2.2 Definitions of Terms Specific to This Standard:
testing. More specifically, this guide is intended to provide for
2.2.1 chromosome mutation—a mutation resulting from a
researchers the accomplishment of the following goals:
structural change to a chromosome involving the gain, loss, or
1.1.1 Provide an understanding of the critical procedures
relocation of chromosome segments. Chromosome mutations
(steps)intheperformanceofthismammaliancellmutagenicity
can be either intrachromosomal or interchromosomal.
test.
2.2.2 relative suspension growth (RSG)—used to measure
1.1.2 Provide generalized criteria by which researchers can
the cytotoxicity of a given treatment based on the growth of
evaluate if they are properly performing, utilizing, and inter-
cells in suspension culture relative to the untreated or solvent
preting this assay.
control(s). RSG is calculated according to the method of Clive
1.1.3 Provide criteria by which individuals responsible for
2
and Spector (1).
evaluating MLA data can determine if the experiments have
2.2.3 relative total growth (RTG)—used as a means to
been properly performed and interpreted.
measure the relative toxicity to cells (survival) following
1.1.4 Provide a basis from which new procedures and
treatment in the mouse lymphoma assay. RTG is calculated
developments in testing procedures can be evaluated.
according to the method of Clive and Spector (1) and includes
1.1.5 Provide an understanding of the types of genetic
RSG as well as the ability to form colonies in the clonal phase
damage (that is, gene and chromosome mutation) that may be
of the assay.
detected in this mammalian cell mutagenicity test.
2.3 Symbols:
2. Terminology
2.3.1 BrUdR—5-bromo-28-deoxyuridine.
2.3.2 BUdR—bromouracil deoxyriboside.
2.1 Definitions:
2.3.3 CAS—chemical abstract service.
2.3.4 DMSO—dimethylsulfoxide.
2.3.5 MLA—mouse lymphoma assay.
1
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.
2
Current edition approved Sept. 10, 2003. Published September 2003. Originally Theboldfacenumbersinparenthesesrefertothelistofreferencesattheendof
approved in 1989. Last previous edition approved in 1997 as E1280–97. 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 (2003)
2.3.6 NADP—nicotinamide-adenine dinucleotide phos- Baltimore Biological Laboratories (BBL); see Ref. (8)) and
phate. often with additional serum. Each investigator should deter-
2.3.7 TFT—trifluorothymidine. mine serum and agar concentrations that yield the best cloning
2.3.8 THMG—thymidine + hypoxanthine + methotrexate + conditionsintheirlaboratory.Seereferencesin4.1foragarand
glycine. serumconcentrationsastheyvarybetweenlaboratories.
...