研究用肿瘤细胞系的质量控制和要求.ppt

1、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 45/WISH; 59/Chang liver; 470/Hep-2; 556/KB; 2004(Buehring

2、), 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-

3、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-transformed amniotic epithelial cells”) in Mol. Pharmacol. 69, 796(2006), Endocrinology 147, 2490 (2006) and J. Biol

4、. Chem. 278, 31731 (2003) Chang liver (described as “normal liver cells”) in Oncogene 28, 3526 (2009), Proteomics 14, 2885(2008) and J. Biol. Chem. 279, 28106 (2000) HEp-2 (described as “laryngeal cancer”) in Investig. New Drugs 26, 111118 (2008),Carcinogenesis 29, 1519 (2008) and J. Biol. Chem. 283

5、, 36272 (2008) KB (described as “oral cancer”) in Biochem. Pharmacol. 73,19011909 (2007), Clin. Cancer Res. 14,8161(2008) and J. Biol. Chem. 280, 23829 (2005) HeLa, Int-407 and HEp-2 cells were used as three distinct cell lines in the same study in CancerRes. 69, 632 (2009),Ecv-304,“人脐静脉内皮细胞”,日本实验室建

6、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? Donot know? No other suitable cellular model Expensieve, can not afford, have to compromise,Ecv-304/T24,对细胞质量的要求,2007.10,Goslar, Germany，11ICCC Scientist from ATCC proposed:(1)funding agencies,g

7、ov. or private require cell line authentication as a condition for the award of grant and contracts funds. (2)key scientific J as a condition for publication. (3) conference, workshop and/or training.(4)lab heads be encouraged to ensure QC measures. Authentic, Well characterized The problem of Sub-c

8、ulturing,NIH NOTICE,2007.12, Require cell anthentication For grant For paper,杂志及细胞使用者的要求Examples 1,Thp-1, 2009-1-5 cell line authentication, characterization and freedom from contamination This is an important issue, to which authors should pay particular attention. It is the authors responsibility

9、to ensure that cell lines used in any study (and the majority do involve them) are certified as being the designated type, that they have been checked to ensure they are free of contamination (e.g. mycoplasma), and that they have been used from young stock. These are the three criteria upon which go

10、od practice is based, as set out in the UKCCC Guidelines on the use of cell lines ( http:/ukcccr.icnet.uk). Attention should be paid to this in the Material and Methods section, and authors omitting to include evidence of good practice will find that their articles may be delayed until such guarante

11、es are given.,Example 2, Reporting on Cell Line Use in AACR Journals,NEW! EFFECTIVE JULY 1, 2009 AACR strongly encourages the authentication of cell lines used in the research reported in its journals. If cell lines were used in the research, a statement addressing the following points must be inclu

12、ded in the Materials and Methods section: 1. From where and when the cells were obtained 2. Whether the cell lines have been tested and authenticated 3. The method by which the cells were tested 4. How and when the cells were last tested,AACR Journals,Cancer Research Clinical Cancer Research Cancer Epidemiology, Biomarkers 14951509. Cell Line Verification Test Recommendations, ATCC Technical Bulletin No.8 (2008). Cell Line Authentication Methods Cabrera CM et al., Identity Test: Determination of Cell