研究用肿瘤细胞系的质量控制和要求.ppt

研究用肿瘤细胞系的质量控制和要求,中国医学科学院基础医学研究所刘玉琴,SomeFactsabouttheuseofculturedcells-Nobelprize,2001，Cellcycle2002,apoptosis2006,RNAInterference2007,homologousrecombinationandgenetargeting2008,HPVcervicalca,HIV-aids2009,telomere45/WISH;59/Changliver;470/Hep-2;556/KB;2004(Buehring),pubmedsearch220publicationsinvolved,32%useHeLacells,33%testedforauthenticity,35%getcellsfromotherlabs.NCIpanelof60celllineswereusedforanticancerdrugevaluation.AmongthemADR-RESisacutallyOVCAR-8.Ithadbeenusedinabout300papers.,HeLacervicalcancercells,Int-407(describedas“non-transformedintestinalepithelialcells”)inBr.J.Cancer101,1596(2009),EMBOJ.22,5003(2003)andJ.Biol.Chem.280,13538(2005)WISH(describedas“non-transformedamnioticepithelialcells”)inMol.Pharmacol.69,796(2006),Endocrinology147,2490(2006)andJ.Biol.Chem.278,31731(2003)Changliver(describedas“normallivercells”)inOncogene28,3526(2009),Proteomics14,2885(2008)andJ.Biol.Chem.279,28106(2000)HEp-2(describedas“laryngealcancer”)inInvestig.NewDrugs26,111118(2008),Carcinogenesis29,1519(2008)andJ.Biol.Chem.283,36272(2008)KB(describedas“oralcancer”)inBiochem.Pharmacol.73,19011909(2007),Clin.CancerRes.14,8161(2008)andJ.Biol.Chem.280,23829(2005)HeLa,Int-407andHEp-2cellswereusedasthreedistinctcelllinesinthesamestudyinCancerRes.69,632(2009),Ecv-304,“人脐静脉内皮细胞”,日本实验室建立，ATCC、DSMZ等机构有保藏，在19992000年，经STR检测，证明为T24(膀胱癌)，各自网站有公布。在实验室间广泛传播，2006y,全世界600余论文，利用该细胞进行研究，2/3在中国。Why?Donotknow?NoothersuitablecellularmodelExpensieve,cannotafford,havetocompromise,Ecv-304/T24,对细胞质量的要求,2007.10,Goslar,Germany，11ICCCScientistfromATCCproposed:(1)fundingagencies,gov.orprivaterequirecelllineauthenticationasaconditionfortheawardofgrantandcontractsfunds.(2)keyscientificJasaconditionforpublication.(3)conference,workshopand/ortraining.(4)labheadsbeencouragedtoensureQCmeasures.Authentic,WellcharacterizedTheproblemofSub-culturing,NIHNOTICE,2007.12,RequirecellanthenticationForgrantForpaper,杂志及细胞使用者的要求Examples1,Thp-1,2009-1-5celllineauthentication,characterizationandfreedomfromcontaminationThisisanimportantissue,towhichauthorsshouldpayparticularattention.Itistheauthorsresponsibilitytoensurethatcelllinesusedinanystudy(andthemajoritydoinvolvethem)arecertifiedasbeingthedesignatedtype,thattheyhavebeencheckedtoensuretheyarefreeofcontamination(e.g.mycoplasma),andthattheyhavebeenusedfromyoungstock.Thesearethethreecriteriauponwhichgoodpracticeisbased,assetoutintheUKCCCGuidelinesontheuseofcelllines(http:/ukcccr.icnet.uk).AttentionshouldbepaidtothisintheMaterialandMethodssection,andauthorsomittingtoincludeevidenceofgoodpracticewillfindthattheirarticlesmaybedelayeduntilsuchguaranteesaregiven.,Example2,ReportingonCellLineUseinAACRJournals,NEW!EFFECTIVEJULY1,2009AACRstronglyencouragestheauthenticationofcelllinesusedintheresearchreportedinitsjournals.Ifcelllineswereusedintheresearch,astatementaddressingthefollowingpointsmustbeincludedintheMaterialsandMethodssection:1.Fromwhereandwhenthecellswereobtained2.Whetherthecelllineshavebeentestedandauthenticated3.Themethodbywhichthecellsweretested4.Howandwhenthecellswerelasttested,AACRJournals,CancerResearchClinicalCancerResearchCancerEpidemiology,Biomarkers14951509.CellLineVerificationTestRecommendations,ATCCTechnicalBulletinNo.8(2008).CellLineAuthenticationMethodsCabreraCMetal.,IdentityTest:DeterminationofCellLin