Detection algorithm for the validation of human cell lines

Cell lines are an important tool in understanding all aspects of cancer growth, development, metastasis and tumor cell death. There has been a dramatic increase in the number of cell lines and diversity of the cancers they represent; however, misidentification and cross‐contamination of cell lines can lead to erroneous conclusions. One method that has gained favor for authenticating cell lines is the use of short tandem repeats (STR) to generate a unique DNA profile. The challenge in validating cell lines is the requirement to compare the large number of existing STR profiles against cell lines of interest, particularly when considering that the profiles of many cell lines have drifted over time and original samples are not available. We report here methods that analyze the variations and the proportional changes extracted from tetra‐nucleotide repeat regions in the STR analysis. This technique allows a paired match between a target cell line and a reference database of cell lines to find cell lines that match within a user designated percentage cut‐off quality matrix. Our method accounts for DNA instability and can suggest whether the target cell lines are misidentified or unstable.

[1]  Wilhelm G Dirks,et al.  Cell line cross‐contamination initiative: An interactive reference database of STR profiles covering common cancer cell lines , 2010, International journal of cancer.

[2]  J A Thomson,et al.  Short tandem repeat profiling provides an international reference standard for human cell lines , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[3]  A. Monks,et al.  Characterization of MLH1 and MSH2 DNA mismatch repair proteins in cell lines of the NCI anticancer drug screen , 2000, Cancer Chemotherapy and Pharmacology.

[4]  Y. Matsuo,et al.  False leukemia–lymphoma cell lines: an update on over 500 cell lines , 2003, Leukemia.

[5]  Amanda Capes-Davis,et al.  Recommendation of short tandem repeat profiling for authenticating human cell lines, stem cells, and tissues , 2010, In Vitro Cellular & Developmental Biology - Animal.

[6]  W. Nelson-Rees,et al.  Inter- and intraspecies contamination of human breast tumor cell lines HBC and BrCa5 and other cell cultures , 1977, Science.

[7]  Jeffrey S. Morris,et al.  The critical role of 15-lipoxygenase-1 in colorectal epithelial cell terminal differentiation and tumorigenesis. , 2005, Cancer research.

[8]  R. Costello,et al.  Impostor Cell Lines , 2006, The Laryngoscope.

[9]  S. Narayan Curcumin, A Multi-Functional Chemopreventive Agent, Blocks Growth of Colon Cancer Cells by Targeting β-Catenin-Mediated Transactivation and Cell–Cell Adhesion Pathways , 2003, Journal of Molecular Histology.

[10]  H. Drexler,et al.  False human hematopoietic cell lines: cross-contaminations and misinterpretations , 1999, Leukemia.

[11]  H. Drexler,et al.  Widespread intraspecies cross‐contamination of human tumor cell lines arising at source , 1999, International journal of cancer.

[12]  Stephen Frosh,et al.  Identity crisis , 2015, Science.

[13]  Roland M. Nardone,et al.  Eradication of cross-contaminated cell lines: A call for action , 2007, Cell Biology and Toxicology.

[14]  Paolo Romano,et al.  Cell Line Data Base: structure and recent improvements towards molecular authentication of human cell lines , 2008, Nucleic Acids Res..

[15]  M. Peinado,et al.  Cytogenetic characterization of two colon cell lines by using conventional G-banding, comparative genomic hybridization, and whole chromosome painting. , 2000, Cancer genetics and cytogenetics.

[16]  Wilhelm G Dirks,et al.  One falsehood leads easily to another , 2008, International journal of cancer.

[17]  J. Bubeník Cross-contamination of cell lines in culture. , 2000, Folia biologica.

[18]  M. Lacroix,et al.  Persistent use of “false” cell lines , 2008, International journal of cancer.

[19]  Stefan Schmidt,et al.  Cancer cell line identification by short tandem repeat profiling: power and limitations , 2005, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[20]  Wilhelm G Dirks,et al.  Online verification of human cell line identity by STR DNA typing. , 2011, Methods in molecular biology.

[21]  Sudhir Varma,et al.  DNA fingerprinting of the NCI-60 cell line panel , 2009, Molecular Cancer Therapeutics.

[22]  Paula Chaves,et al.  Verification and unmasking of widely used human esophageal adenocarcinoma cell lines. , 2010, Journal of the National Cancer Institute.

[23]  G. Vogel Cell biology. To scientists' dismay, mixed-up cell lines strike again. , 2010, Science.