武汉普诺赛生命科技有限公司
当前位置:首页 >> 细胞资源 >> 细胞STR鉴定

细胞资源Cell Resources

联系我们CONTACT US

细胞STR鉴定简介:

细胞STR鉴定应用于生物医学研究领域的哺乳动物细胞被错误鉴定和交叉污染的问题,一直是一个普遍存在的突出问题。

据统计,国外实验室有20%左右的细胞株被错误鉴定和交叉污染,NIH和ATCC近两年都对此发出呼吁,要求研究者对细胞进行鉴定。近年来,大量研究表明STR基因分型方法是进行细胞交叉污染和性质鉴定的最有效和准确的方法之一,STR基因分型应用于细胞鉴定已被ATCC等机构强烈推荐。美国的ATCC细胞库、德国的DSMZ细胞库以及日本的JCRB细胞库等为STR分型提供了各细胞株的数据供比对。

STR(Short Tandem Repeat,短串联重复序列)基因位点由长度为3-7个碱基对的短串连重复序列组成,这些重复序列广泛存在于人类基因组中,可作为高度多态性标记,被称为细胞的DNA指纹,其可通过PCR(聚合酶链式反应)来检测。STR基因座位上的等位基因可通过扩增区域内重复序列的拷贝数的不同来区分,在毛细管电泳分离之后可通过荧光检测来识别。随后通过一定的计算方法,即可根据所得的STR分型结果与专业的细胞STR数据库比对从而推算出样品所属的细胞系或可能的交叉污染的细胞系名称。


细胞STR鉴定服务流程:

1515554798798411.png


细胞STR鉴定样本要求:

1515556125177678.png

细胞系错误鉴定所造成的后果:

★ 细胞系的损失;

★ 时间和金钱的损失;

★ 在公众领域(文献、专利等)提供错误信息;

★ 造成实验结果不一致或不可重复;

★ ……


何时需做细胞STR鉴定?

1、发表文章或申请课题经费前;

2、使用细胞进行临床治疗(试验)前;

3、准备冻存保种或已冻存多年的细胞;

4、一个涉及到细胞试验项目开始/结束时;

5、新得到的细胞或实验室传5代以上的细胞;

6、细胞系表现不稳定或结果与预期差别较大。


已开始要求细胞鉴定的期刊:

★ Nature

★ BioTechniques

★ Cancer Research

★ Cancer Discovery

★ Clinical Cancer Research

★ Molecular Cancer Research

★ Cancer Prevention Research

★ International Journal of Cancer

★ Molecular Cancer Therapeutics

★ Cell Biochemistry and Biophysics

★ Cancer Epidemiology, Biomarkers & Prevention

★ In Vitro Cellular & Developmental Biology - Animal

★ ......


人源细胞STR位点信息:

1515554785615354.png


为进一步提高鉴定的分辨率,除了包含ATCC检测的8个STR和1个性别决定位点Amelogenin外,另新增了12个高度多态性位点,共检测21个STR位点。


参考文献:

1. Chatterjee, R. (2007) Cell biology. Cases of mistaken identity. Science 315, 928-31.

2. Ruiz Bravo, N. and Gottesman, M. (2007) Notice regarding authentication of cultured cell lines.

3. Yoshino, K. et al. (2006) Essential role for gene profiling analysis in the authentication of human cell lines. Human Cell 19, 43-8.

4. Szibor, R. et al. (2003) Cell line DNA typing in forensic genetics—the necessity of reliable standards. Forensic Sci. Int. 138, 37-43.

5. Dirks, W.G. et al. (2005) Short tandem repeat DNA typing provides an international reference standard for authentication of human cell lines. ALTEX 22, 103-9.

6. Masters, J.R. et al. (2001) Short tandem repeat profiling provides an international reference standard for human cell lines. Proc. Natl. Acad. Sci. USA 98, 8012-7.

7. (2001) Verify cell line identity with DNA profiling. ATCC Connection: Newsletter of The American Type Culture Collection 21, 1-2.

8. Krenke, B. et al. (2002) Validation of a 16-locus fluorescent multiplex system. J. Forensic Sci. 47, 773-85.

9. Levinson, G. and Gutman, G.A. (1987) Slipped-strand mispairing: A major mechanism for DNA sequence evolution. Mol. Biol. Evol. 4, 203-21.

10. Schlotterer, C. and Tautz, D. (1992) Slippage synthesis of simple sequence DNA. Nucleic Acids Res. 20, 211-5.

11. Smith, J.R. et al. (1995) Approach to genotyping errors caused by nontemplated nucleotide addition by Taq DNA polymerase. Genome Res. 5, 312-7.

12. Magnuson, V.L. et al. (1996) Substrate nucleotide-determined non-templated addition of adenine by Taq DNA polymerase: Implications for PCR-based genotyping. BioTechniques 21, 700-9.

13. Walsh, P.S., Fildes, N.J. and Reynolds, R. (1996) Sequence analysis and characterization of stutter products at the tetranucleotide repeat locus vWA. Nucleic Acids Res. 24, 2807-12.

14. Moller, A., Meyer, E. and Brinkmann, B. (1994) Different types of structural variation in STRs: HumFES/FPS, HumVWA and HumD21S11. Int. J. Leg. Med. 106, 319-23.

15. Brinkmann, B., Moller A. and Wiegand, P. (1995) Structure of new mutations in 2 STR systems. Int. J. Leg. Med. 107, 201-3.

16. Griffiths, R. et al. (1998) New reference allelic ladders to improve allelic designation in a multiplex STR system. Int. J. Legal Med. 111, 267-72.

17. Bär, W. et al. (1997) DNA recommendations: Further report of the DNA Commission of the ISFH regarding the use of short tandem repeat systems. Int. J. Legal Med. 110, 175-6.

18. Gill, P. et al. (1997) Considerations from the European DNA Profiling Group (EDNAP) concerning STR nomenclature. Forensic Sci. Int. 87, 185-92.