13EYO1007550遗传毒性研究小鼠骨髓微核实验500ml600ml400KIMAX300200NO:14000
遗传毒性研究 小鼠骨髓微核实验
微核试验（简称：MNT)发展历史1959年Evans等蚕豆根端细胞电脑辐射暴露抗肿瘤药三亚胺醒给药中国黄金地鼠1970年Boller和Schmid，正式命名MNT1976年人外周血培养淋巴细胞进行MNTCountryman, Heddle妊娠雌鼠给药，胎仔小鼠的肝细胞和1979年Cole，King和Wild等外周血出现微核1980年MacGregor等小鼠外周血检测MN方法1981年和Lahdetie,Tates等精母细胞MNT，预测生殖毒性1983年1983年荧光染料AO特异性染色引入MNTHayashi,MacGregor等
1959年 Evans等 蚕豆根端细胞电脑辐射暴露 1970年 Boller和Schmid 抗肿瘤药三亚胺醌给药中国黄金地鼠 ，正式命名MNT 1976年 Countryman，Heddle 人外周血培养淋巴细胞进行MNT 1979年 Cole，King和Wild等 妊娠雌鼠给药，胎仔小鼠的肝细胞和 外周血出现微核 1980年 MacGregor等 小鼠外周血检测MN方法 1981年和 1983年 Lahdetie,Tates等 精母细胞MNT，预测生殖毒性 1983年 Hayashi, MacGregor等 荧光染料AO特异性染色引入MNT 微核试验（简称：MNT)发展历史
TG 474OECD/OCDE26 September 2014指导原则OECDGUIDELINEFORTHETESTINGOFCHEMICALS474MAMIMALIANERYTHROCYTEMICRONUCLEUSTESTAdopted:INTRODUCTION29July2016Ds OECD.Gasadketetm1983.In199revised veradotedLbaiedoniefiepeogresdeto thatdate.Thismodified version oftheTest Cmidelineeflectnof.thedamdinhaFoitcabes ofTetCuidliOECD GUIDELINE FOR THE TESTING OF CHEMICALSMammalianErvthrocyteMicronucleusTestINTRODUCTION1.The OECD Guidelines for the Testing of Chemicals are periodically reviewed in the light ofscientific progress, changing regulatory needs and animal welfare considerations. The original TestGuideline474was adopted in 19s3.In1997.a revised version was adopted,based on scientific progressmade to that date.This modified version of the TestGuidelinereflects scientific knowledge from morethan thirty years of experience with this assay and the interpretation of the data, and in particular theadvances in automated scoring technologies and the potential for integrating or combining this test withother general toxicity orgenotoxicity studies.This Test Guideline is part of a series of Test Guidelines ongenetic toxicology.A document that provides succinct information on genetic toxicology testing and an
指导原则
Office of FoodAdditive SafetyRedbook2000Toxicological Principlesforthe SafetyAssessment of Food IngredientsJuly2000:UpdatedOctober,2001Niemher2003&Anrit200IV.C.1.d. Mammalian ErythrocyteIntrdMicronucleusTestThis guidance represents the Foodthinking on thistopic.It does notcand does not operate tobindFDAI.Introductionapproach if such an approach satisfandregulations.Ifyouwanttodisstaffresponsibleforimplementing301-436-1200).Contact informatiMicronuclei are cytoplasmic chromatin-containingbodiesformedwhen acentricForquestionsregardingtheuseorchromosomefragments or chromosomeslagduringanaphase andfailtobecomeprinted copies of individual chapteincorporated into daughter cell nuclei during cell division.Because geneticdamageAdditiveSafety (OFAS)thatresults inchromosomebreaks,structurallyabnormal chromosomes,or spindleabnormalities leads tomicronucleus formation,the incidenceofmicronuclei servesas an index ofthesetypes of damage. It has been established that essentially allagents that causedouble strand chromosomebreaks (clastogens)inducemicronucleiBecause enumeration ofmicronucleiismuchfaster and less technically demandingthanis scoring ofchromosomal aberrations,and becausemicronucleiarisefromtwo importanttypes of geneticdamage(clastogenesis and spindledisruption),themicronucleusassayhasbeen widelyused toscreenforchemicals thatcausethesetypes of damage
4.RECOMMENDATIONSFORINVIVOTESTS4.1TestsfortheDetectionofChromosomeDamageInVioEitherthe analysis of chromosomal aberrationsorthe measurement of micronucleatedpolychromaticerythrocytes inbonemarrowcells invivois consideredappropriateforthedetection of clastogens.Both rats and mice are considered appropriate for use in thebone marrow micronucleus test.Micronuclei can also be measured in immature (e.g.,polychromatic) erythrocytes in peripheral blood in the mouse, orin the newly formedreticulocytes in rat blood (Note 3).Likewise,immature erythrocytes can be used fromanyotherspecieswhichhasshownanadequatesensitivitytodetectclastogens/aneuploidy inducers in bone marrow or peripheral blood (Note 3). Systemsfor automated analysis (image analysis andflow cytometry)can beusedif appropriatelyvalidated (OECD.1997:Hayashiet al..2000:2007).Chromosomal aberrationscanalsobeanalyzedinperipherallymphocytesculturedfromtreatedrodents (Notell).4.2OtherInViuoGenotoxicityTestsICHHARMONISEDTRIPARTITEGUIDELINEThe same in vivo tests described as the secondcan be used as follow-up tests to develop weigvitro or invivo assays (Notes 1l and 12).Whilknowledge of the mechanism can help guide theGUIDANCEONGENOTOXICITYTESTINGANDchromosomal aberrations or of gene mutationsDATAINTERPRETATIONFORPHARMACEUTICALSINTENDEDFORHUMANUSEstandardmethodsinmosttissues.Althoughnin rodents, this entails prolonged treatmentS2(R1)expression,fixation andaccumulation,especiall12).Thus the second in vivo assay will oftensurrogate. Assays with the mostpublished exy
实验原理是染色体或染色单体的无着丝点断片或纺锤丝受损伤而丢失的整个染色体。>可检出DNA断裂剂和非整倍体诱变剂6
6 是染色体或染色单体的无着丝点断片或纺锤丝受 损伤而丢失的整个染色体。 可检出DNA断裂剂和非整倍体诱变剂。 实验原理
微核细胞形成过程骨髓外周血染色体断片药物处理网织红细胞正常红细胞I分裂成熟0正染红细胞分裂中期母红细胞分裂末期微核红细胞微核网织红细胞10h10h10h骨髓中检测到微核在18-30h（给药2次，通常在18-24h采样外周血液中检测到微核要>30h
微核细胞形成过程 10h 10h 10h 母红细胞 分裂中期 分裂末期 分裂 正常红细胞 微核红细胞 药物处理 染色体断片 网织红细胞 微核网织红细胞 成熟 正染红细胞 骨髓 外周血 骨髓中检测到微核在18-30h（给药2次，通常在18-24h采样） 外周血液中检测到微核要>30h
NucleusMicronucleusRats/micedosedwithcompound,3Micronucleimaybeformedbylossofwholechromosomedoses,6animals/group.Animalsduringdivisionorbychromosomebreakage.Theerythrocyte'ssacrificed24or48hourslaternucleusisextruded leavinganymicronucleibehindMicronucleusBonemarrowcellsspreadontoFemursremovedslides.Slidesfixedand stainedand bone marrow2000cellsanalysedperanimal,numberaspirated(acridine orange)ofmicronucleatedimmatureerythrocytesscored
Rats/mice dosed with compound, 3 doses, 6 animals / group. Animals sacrificed 24 or 48 hours later Micronuclei may be formed by loss of whole chromosome during division or by chromosome breakage. The erythrocyte’s nucleus is extruded leaving any micronuclei behind Nucleus Micronucleus Femurs removed and bone marrow aspirated Bone marrow cells spread onto slides. Slides fixed and stained (acridine orange) 2000 cells analysed per animal, number of micronucleated immature erythrocytes scored Micronucleus
Rodentbone-marrowmicronucleustestNCE
Rodent bone-marrow micronucleus test
5012吖啶橙染色后显示为肥大颗粒
吖啶橙染色后显示为肥大颗粒