ASTM_E_1202_-_87_2003.pdf

Designation:E 1202 87(Reapproved 2003)Standard Guide forDevelopment of Micronucleus Assay Standards1This standard is issued under the fixed designation E 1202;the number immediately following the designation indicates the year oforiginal adoption or,in the case of revision,the year of last revision.A number in parentheses indicates the year of last reapproval.Asuperscript epsilon(e)indicates an editorial change since the last revision or reapproval.1.Scope1.1 This guide covers minimal criteria which should be metby a micronucleus assay system prior to the development of anASTM Standard or Guide for the conduct of that assay.1.2 This standard does not purport to address all of thesafety concerns,if any,associated with its use.It is theresponsibility of the user of this standard to establish appro-priate safety and health practices and determine the applica-bility of regulatory limitations prior to use.2.Significance and Use2.1 Micronucleus assays for genetic damage have beendeveloped in many types of eucaryotic cells,both in vitro andin vivo.The occurrence of micronuclei is indicative of chro-mosomal damage or mitotic spindle dysfunction.3.Criteria3.1 Biology:3.1.1 The biology of the system should be well understoodin terms of(a)cell cycle,(b)metabolic capabilities,(c)cultureor growth conditions,and(d)other factors of importance inmaintaining a reproducible experimental situation.Thereshould be evidence that micronuclei arise from chromosomalaberrations or chromosome loss or both,and not apoptosis orany other mechanism.3.2 Time Response:3.2.1 The“expression time”for micronuclei should becharacterized for(a)direct-acting genotoxins and(b)genotox-ins requiring metabolic activation.3.3 Dose Response:3.3.1 The dose response curves for several classes ofgenotoxins,over a dose range including both toxic andnontoxic doses,should be known.A rational method fordetermining the upper and lower doses to be tested should beavailable.3.4 Spontaneous Frequency:3.4.1 The spontaneous frequency of micronuclei should bewell characterized and should be stable under the test condi-tions employed.Major factors affecting the spontaneous inci-dence of micronuclei should be defined.3.5 Statistics:3.5.1 The following statistical criteria should be met:3.5.1.1 There should be sufficient data to define the majorsources of experimental variability(for example,slide to slide,animal to animal),in order to permit rational experimentaldesign,3.5.1.2 Appropriate statistical methods for analyzing thedata should be available,3.5.1.3 Sufficient data and adequate statistical methodsshould be available to permit determination of the sample sizesrequired for adequate statistical power,and3.5.1.4 The quantitative reproducibility of experimental re-sults between and within experiments should be known.3.6 Transportability:3.6.1 There should be sufficient experience with the systemin order to know how well the characteristics of the assay aremaintained in different laboratories.It should be knownwhether observer-dependent effects,such as scoring andsample preparation,have been sufficiently controlled amonglaboratories to ensure uniform interpretation of test data.Theinfluence of factors,such as source of test organism andmaterials,on experimental outcome should be known.Awritten description of the techniques required for the conductof the assay that is adequate to permit a new laboratory withnormal experience in genetic toxicology testing to carry out theassay should be available.1This guide is under the jurisdiction of ASTM Committee F04 on Medical andSurgical Materials and Devices and is the direct responsibility of SubcommitteeF04.16 on Biocompatibility Test Methods.Current edition approved Sept.10,2003.Published September 2003.Originallyapproved in 1987.Last previous edition approved in 1998 as E 1202 87(1998).1Copyright ASTM International,100 Barr Harbor Drive,PO Box C700,West Conshohocken,PA 19428-2959,United States.ASTM International takes no position respecting the validity of any patent rights asserted in connection with any item mentionedin this standard.Users of this standard are expressly advised that determination of the validity of any such patent rights,and the riskof infringement of such rights,are entirely their own responsibility.This standard is subject to revision at any time by the responsible technical committee and must be reviewed every five years andif not revised,either reapproved or withdrawn.Your comments are invited either for revision of this standard or for additional standardsand should be addressed to ASTM International Headquarters.Your comments will receive careful consideration at a meeting of theresponsible technical committee,which you may attend.If you feel that your comments have not received a fair hearing you shouldmake your views known to the ASTM Committee on Standards,at the address shown below.This standard is copyrighted by ASTM International,100 Barr Harbor Drive,PO Box C700,West Conshohocken,PA 19428-29