The Role of Cell Line Authentication in Today’s Biomedical Research World

Introduction

Inadvertent cell line contamination is a serious concern for many researchers. Failure to monitor cell lines for contamination may result in compromised data. Current estimates are that 18% to 36% of all active cell lines are either cross-contaminated or misidentified.1 This level of concern has led a number of journals to actively encourage cell line authentication prior to submission for publication. Cell line authentication testing is a good quality control step for any laboratory utilizing human cell lines and/or stem cell lines. If you want to set your research on a strong foundation, please join us for this week’s Ask the Expert – “The Role of Cell Line Authentication in Today’s Biomedical Research World,” and ask any questions you have about Cell Line Authentication.

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Question 1

I perform Karyotyping and isoenzyme analysis in my lab, why is STR DNA profiling a better technology than other methods for authentication?

STR DNA profiling is a method of human identification that allows for determination that the cell line was derived from a specific human – no two humans have the same STR DNA profile except for identical twins. Typically, karyotyping and isoenzyme analysis do not distinguish individual humans. Total: 0 0 0 0

Question 2

How does cell line misidentification and cross-contamination happen in the lab?

Unfortunately, mistakes can happen in even the strictest of labs. The infamous HeLa cells, which are extremely aggressive and fast growing, are known for being able to contaminate other cultures via aerosol droplets. A simple mislabeling of a flask could cause misidentification of the cell line in use. Not following stringent cell culture rules is … Continued

Question 3

What journals require cell line authentication testing prior to publication?

There are currently several well-known journal publishers that have guidelines for cell line authentication testing prior to submission including (see instructions to authors for all journals): 1. American Association for Cancer Research (AACR) Journals, which includes – Cancer Discovery, Cancer Epidemiology, Biomarkers and Prevention, Cancer Prevention Research, Cancer Research, Clinical Cancer Research, Molecular Cancer Research, … Continued

Question 4

How do you recommend companies implement cell line authentication? At what point in the process would you get your lines tested and what are the issues that can happen if you choose not to test your cell lines? Thanks

A) Companies looking to implement a cell line authentication program need to decide if they wish to perform the testing internally or reference the samples to a laboratory specializing in cell line authentication. To test internally, the technology currently recommended by ANSI-ATCC standards is STR DNA analysis. This would require purchasing a genetic analyzer, thermal … Continued

Question 5

What can the STR DNA profile tell me about a cell line?

The STR DNA profile can determine if there is no contamination or alteration of the cultured cell line as compared to the original cells. The results may be able to identify a contaminant and also measure instability or other changes in the cell line. There are several mathematical measures that can assist in the probability … Continued

Question 6

Are there any regulatory requirements to perform cell line authentication?

There are no specific regulations for cell line testing. See below for information on characterizing cell lines. US FDA – “Points to consider in the characterization of cell lines used to produce biologicals. Department of Health and Human Services, Center for Biologics Evaluation and Research, 1993”. For information on cell line authentication requirements for journal … Continued