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The Prostate Cancer Foundation Unveils Major Initiative to Improve Accuracy of Laboratory Cancer Studies

Best Practices in Cell Line Authentication

Howard Soule, PhD

Executive Vice President and Chief Science Officer, PCF

November 21, 2013 - A cell line is a permanently maintained culture of cells in the laboratory that is used to conduct experiments. Cell lines are commonly used to study cancer. Cancer cell lines date back to the 1950s, when the first line was established from a woman, Henrietta Lacks, with cervical cancer. In the decades since, approximately 1,000 cancer cell lines have become essential tools to study cancer cell biology and to test drugs. Indeed, almost every anti-cancer drug now in use gained early traction through testing on cancer cell lines.1

Prostate cancer cell lines are very important tools in the fight against this disease. However, there are problems with cell lines that need to be addressed. A study published in the Proceedings of the National Academy of Sciences in 2011 that looked at 60 established cancer cell lines found that the cell lines studied more closely resembled other cell lines than the cancer type they were supposed to model.1,2 Another study from the Journal of Endocrinology and Metabolism used genetic analysis techniques (short tandem repeat analysis and single nucleotide polymorphism array analysis) to evaluate 40 thyroid cancer-derived cell lines that had been widely used to study thyroid cancer for 20 years.3 The researchers found the thyroid cell lines to be contaminated with colon and skin cancer cells—only 23 of the 40 lines studied proved to be “pure” for their unique cell line characteristic.

Cell Line Misidentification a Widespread Problem in Labs

The problem of contaminated cancer cell lines is widespread. Over one-third of human cell lines used for biomedical research are either misidentified or contaminated.3 Adding to the problem: About one-third of continuous cell line cultures in academic institutions are contaminated with a bacteria known as mycoplasma.4 Such contamination can affect study results. Soule pointed to a list of ten serious consequences of cell line contamination with mycoplasma bacteria.

Source: Pall Corporation

The Prostate Cancer Foundation (PCF) has put out a mission statement to help resolve this problem, and at the 20th Annual Scientific Retreat Dr. Soule gave a presentation outlining this mission statement. The Prostate Cancer Foundation has undertaken a Cell Line Authentication Initiative (CLAI). This calls for widespread implementation of best practices for cell line authentication. Today rapid and accurate DNA profiling techniques such as those mentioned above are available, and when used to verify cell line integrity provide a solution to the problem of misidentified and contamination.5 Scientists writing in the Journal of the National Cancer Institute this year said: Cancer cell lines have been marked by both success and failure. Cell line misidentification arose as the first problem, which should now be minimized by the launch of a standardized protocol for the authentication of human cell lines using short tandem repeat profiling.6

cell line authentication


The Prostate Cancer Cell Line Authentication Initiative

Dr. Soule said the mission of the PCF Cell Line Authentication Initiative (CLAI) is four pronged.

First, the CLAI will raise awareness of the issue of cell line misidentification and pathogen contamination and how cell line impurity can affect research outcomes.

Second, the Prostate Cancer Foundation will determine the extent of cell line misidentification and pathogen contamination of human cell lines used by PCF-funded researchers. To accomplish this, all cell lines with be genetically analyzed and compared to database from the American Type Culture Collection (ATCC), a non-profit organization devoted to characterizing biological samples such as cell lines. In addition, genetic testing for the presence of mycoplasma in cell lines will be done. All future research funded by the PCF will require ongoing adherence to these best practices.

Third, PCF will help build awareness of the impact of not implementing ongoing, routine cell line authentication in laboratories that use cell lines.

Fourth, PCF will encourage scientific journals and funding agencies to require CLAI and pathogen testing for mycoplasma and other relevant infectious organisms. While many journals and funding agencies have strongly recommended that cell lines undergo authentication, the scientific field as a whole has not yet implemented cell line authentication best practices.

During his presentation, Soule announced that DNA Diagnostics Center in Cincinnati, a company that does paternity testing and other genetic analysis would offer to PCF investigators (for free!) five cell line authentications per investigator, up to a total of 1,000 tests. Researchers can go to for more information on this offer.


  1. Woodbury MA. Will the real cancer cell please stand up? Biotechniques. 2012; (cited; The Clinical Relevance of Cancer Cell Lines JNCI J Natl Cancer Inst (2013) 105 (7): 452-458 first published online February 21, 2013 doi:10.1093/jnci/djt007)
  2. Gillet, J.-P., A. M. Calcagno, S. Varma, M. Marino, L. J. Green, M. I. Vora, C. Patel, J. N. Orina, T. A. Eliseeva, et al. 2011. Redefining the relevance of established cancer cell lines to the study of mechanisms of clinical anti-cancer drug resistance. Proceedings of the National Academy of Sciences 108(46):18708-18713
  3. Schweppe RE, et al. 2008. Deoxyribonucleic acid profiling analysis of 40 human thyroid cancer cell lines reveals cross-contamination resulting in cell line redundancy and misidentification. The Journal of Endocrinology and Metabolism. 93(11):4331-41.
  4. Uphoff, C. C. and H. G. Drexler. 2001. Prevention of mycoplasma contamination in leukemia-lymphoma cell lines. Human Cell. 244:244-247.
  5. Masters, J. (Chair), et al. 2012. Authentication of Human Cell Lines: Standardization of STR Profiling (Designation: ANSI/ATCC ASN-0002-2011). ATCC® Standards Development Organization.
  6. Gillet, JP, et al. The Clinical Relevance of Cancer Cell Lines JNCI J Natl Cancer Inst (2013) 105 (7): 452-458 first published online February 21, 2013 doi:10.1093/jnci/djt007

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