研究用肿瘤细胞系的质量控制和要求课件.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研究用肿瘤细胞系的质量控制和要求

SomeFactsabouttheuseofculturedcells--Nobelprize2001，Cellcycle2002,apoptosis2006,RNAInterference2007,homologousrecombinationandgenetargeting2008,HPV—cervicalca,HIV--aids2009,telomeretelomeraseNobelprizewinner,useinvitroculturedcellsfortheirresearches.研究用肿瘤细胞系的质量控制和要求

现代生命科学研究中培养细胞的使用越来越广泛1969~2004：celluseincreasingsteadilyPapersusecellsincrease2~2.5Xbiology,pharmacologyandmedicine----tumorresearch不可少的研究材料、对象、工具Useofcontaminatedormisidentifiedcellsincrease10X研究用肿瘤细胞系的质量控制和要求

Commonproblemsmicrobiologicalcontaminationwithparticularrespecttomycoplasmainfections(incidenceofca.25%)cross-contaminationbyothercelllinesandmisidentificationormisclassification(incidenceofmorethan15%)acquisitionofgeneticandepigeneticvariationsduetoover-passagingofcelllinesleadingtoageneticdriftofcellculturesandtotheselectionofalteredsubclones.研究用肿瘤细胞系的质量控制和要求

Wherefrom?faultycellculturetechniques:sharedreagents,repeateduseofthesamepipetteduringre-feedingoperationsandmanipulationofmultipleculturesatthesametimewithoutadequateisolationofonecelltypefromanother.theintroductionoffalse,contaminatedorover-passagedcelllinesfromothersourcesintothecellculturelaboratory(35%)Mislabeling,lackofawarenessIntentionalco-cultivationduringpropagationofhumanstemorprimarycellsusingafeederlayerderivedfromanotherspecies研究用肿瘤细胞系的质量控制和要求

细胞交叉污染历史回顾1968,ApparentHeLacellcontaminationofhumanheteroploidcelllines.Nature.217:750-7511974，HeLacesscontaminatingculturesaroundtheworld,Sciences184Contaminatedcellcultures.Nature289:2271981,cross-contaminationofcellsinculture.Sciences.212(4493):226-52研究用肿瘤细胞系的质量控制和要求

细胞交叉污染历史回顾1999,Falsehumanhematopoieticcelllins:cross-cantaminationsandmisinterpretations.Leukemia.13(10):1601-72003,Falseleukmia–lymphomacell