研究用肿瘤细胞系的质量控制和要求

单击此处编辑母版标题样式,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,*,*,*,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,Some Facts about the use of cultured cells-Nobel prize,2001，Cell cycle,2002,apoptosis,2006,RNA Interference,2007,homologous recombination and gene targeting,2008,HPVcervical ca,HIV-aids,2009,telomere&telomerase,Nobel prize winner,use in vitro cultured cells for their researches.,现代生命科学研究中培养细胞的使用越来越广泛,19692004：cell use increasing steadily,Papers use cells increase 22.5X,biology,pharmacology and medicine-tumor research,不可少的研究材料、对象、工具,Use of contaminated or misidentified cells increase 10X,Common problems,microbiological contamination with particular respect to mycoplasma infections(incidence of ca.25%),cross-contamination by other cell lines and misidentification or misclassification(incidence of more than 15%),acquisition of genetic and epigenetic variations due to over-passaging of cell lines leading to a genetic drift of cell cultures and to the selection of altered subclones.,Where from?,faulty cell culture techniques:shared reagents,repeated use of the same pipette during re-feeding operations and manipulation of multiple cultures at the same time without adequate isolation of one cell type from another.,the introduction of false,contaminated or over-passaged cell lines from other sources into the cell culture laboratory(35%),Mislabeling,lack of awareness,Intentional co-cultivation during propagation of human stem or primary cells using a feeder layer derived from another species,细胞交叉污染历史回顾,1968,Apparent HeLa cell contamination of human heteroploid cell lines.Nature.217:750-751,1974，HeLa cess contaminating cultures around the world,Sciences 184:1059,1981,Contaminated cell cultures.Nature 289:227,1981,cross-contamination of cells in culture.Sciences.212(4493):226-52,细胞交叉污染历史回顾,1999,False human hematopoietic cell lins:cross-cantaminations and misint erpretations.Leukemia.13(10):1601-7,2003,False leukmia lymphoma cell lines:an update on over 500 cell lines.Leukemia 17,416-426,2004,cell line cross-comtamination:how aware arein vitro cell dev biol anim.40(7):211-5,2007,Cases of mistaken identity.Sciences 315:928-931,2009,Nature calls for global database of cell line STR profiles,保藏机构公布的错误细胞1,细胞系,种属,细胞类型,污染源,种属,细胞类型,数据来源,207,人,前B白血病,REH,人,前B白血病,DSMZ,2474/90,人,胃癌,HT-29,人,结直肠癌,DSMZ,2957/90,人,胃癌,HT-29,人,结直肠癌,DSMZ,3051/80,人,胃癌,HT-29,人,结直肠癌,DSMZ,ADLC-5M,人,肺癌,HELA/-S3,人,宫颈癌,DSMZ,AV3,人,羊膜,HeLa,人,宫颈癌,ATCC,BCC-1/KMC,人,基底细胞癌,HELA/-S3,人,宫颈癌,DSMZ,BM-1640,人,前列腺癌,DU-145,人,前列腺癌,DSMZ,C16,人,胚胎肺成纤维细胞（MRC-5克隆）,HeLa,人,宫颈癌,ECACC,保藏机构公布的错误细胞2,CHANG Liver,人,胚胎肝上皮,HeLa,人,宫颈癌,ATCC;JCRB,COLO-818,人,黑色素瘤,COLO800,人,黑色素瘤,DSMZ,DAMI,人,巨核细胞,HEL,人,红系白血病,DSMZ,ECV304,人,内皮,T24,人,膀胱癌,ATCC,ECV-304,人,正常内皮,T24,人,膀胱癌,DSMZ,EJ,人,膀胱癌,T24,人,膀胱癌,ATCC;JCRB,EPLC3-2M1,人,肺癌,HELA/-S3,人,宫颈癌,DSMZ;JCRB,EPLC-65,人,肺癌,HELA/-S3,人,宫颈癌,DSMZ,F2-4E5,人,胸腺上皮,SK-HEP-1,人,肝癌,DSMZ,F2-5B6,人,胸腺上皮,SH-HEP-1,人,肝癌,DSMZ,FL,人,羊膜,HeLa,人,宫颈癌,ATCC,GHE,人,星状细胞瘤,T-24,人,膀胱癌,DSMZ,Girardi Heart,猪,成年心脏,HeLa,人,宫颈癌,ATCC,HAG,人,T-24,人,膀胱癌,DSMZ,HEp-2,人,成年喉上皮,HeLa,人,宫颈癌,ATCC,HMV-1,人,黑色素瘤,HeLa-S3,人,宫颈癌,DSMZ,HuL-1,HeLa,人,宫颈癌,JCRB,IMC-2,人,颚骨癌,HeLa-S3,人,宫颈癌,DSMZ,Intestine 407,人,小肠上皮,HeLa,人,宫颈癌,ATCC,J-111,HeLa,人,宫颈癌,JCRB,JOSK-I,人,单核细胞白血病,U-937,人,组织细胞淋巴瘤,DSMZ,JOSK-K,人,单核细胞白血病,U-937,人,组织细胞淋巴瘤,DSMZ,JOSK-M,人,单核细胞白血病,U-937,人,组织细胞淋巴瘤,DSMZ,保藏机构公布的错误细胞3,JOSK-S,人,单核细胞白血病,U-937,人,组织细胞淋巴瘤,DSMZ,JTC-17,HeLa,人,宫颈癌,Yamakage(JCRB),KB,人,HeLa,人,宫颈癌,ATCC;JCRB,KO51,K562,人,髓系白血病,DSMZ;JCRB,KOSC-3,Ca9-22,人,JCRB,L132,人,胚胎肺上皮,HeLa,人,宫颈癌,ATCC,LR10.6,人,前B白血病,NALM-6,人,前B 白血病,DSMZ,MaTu,人,乳腺癌,HELA/-S3,人,宫颈癌,DSMZ,MC-4000,人,乳腺癌,HELA/-S3,人,宫颈癌,DSMZ,MKB-1,人,T细胞白血病,CCRF-CEM,人,T细胞白血病,DSMZ,保藏机构公布的错误细胞4,MKN28,MKN74,人,JCRB,MOLT-15,人,T细胞白血病,CTV-1,人,单核细胞白血病,DSMZ,MT-1,人,乳腺癌,HELA/-S3,人,宫颈癌,DSMZ,NCC16,PHK16-0b,人,JCRB,P1-1A3,人,胸腺上皮,SK-HEP-1,人,肝癌,DSMZ,P1-4D6,人,胸腺上皮,SK-HEP-1,人,肝癌,DSMZ,P39 TSU,人,HL60,人,髓系白血病,JCRB,PBEI,人,前B白血病,NALM-6,人,前B白血病,DSMZ,PSV811,人,成纤维,WI38,人,成纤维,JCRB,RAMAK-1,人,肌肉滑膜,T-24,人,膀胱癌,DSMZ,保藏机构公布的错误细胞5,SBC-2,人,膀胱癌,HELA/-S3,人,宫颈癌,DSMZ,SBC-7,人,膀胱癌,HELA/-S3,人,宫颈癌,DSMZ,SCLC-16H,人,SCLC,SCLC-21/22H,人,SCLC,DSMZ,SCLC-24H,人,SCLC,SCLC-21/22H,人,SCLC,DSMZ,SNB-19,U251MG,人,胶质瘤,ATCC,SPI-801,人,T细胞白血病,K562,人,髓系白血病,DSMZ,SPI-802,人,T细胞白血病,K562,人,髓系白血病,DSMZ,TK-1,人?,U251MG,人,胶质瘤,JCRB,TMH-1,IHH-4,人,JCRB,WISH,人,新生羊膜上皮,HeLa,人,宫颈癌,ATCC,保藏机构公布的错误细胞6,DSMZ 德国细胞库,550 leukemia and lymphoma cell lines submitted,59/395(15%)by originators,23/155(15%)by secondary sources,交叉污染细胞使用的后果,Erroneous results:,Wasted research money,Potential delay in discovery,错误细胞使用,2000-2004：19/Int 407;45/WISH;59/Chang liver;470/Hep-2;556/KB;,2004(Buehring),pubmed search 220publications involved,32%use HeLa cells,33%tested for authenticity,35%get cells from other labs.,NCI panel of 60 cell lines were used for anticancer drug evaluation.Among them ADR-RES is acutally OVCAR-8.It had been used in about 300 papers.,HeLa cervical cancer cells,Int-407(described as“non-transformed intestinal epithelial cells”)in,Br.J.Cancer,101,1596(2009),EMBO J.,22,5003(2003)and,J.Biol.Chem.,280,13538(2005),WISH(described as“non-transfo