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Cell Identity

Lab Academy

According to ICLAC (International Cell Line Authentication Committee) terms of reference, a cell line is considered to be misidentified if its DNA profile (genotype) is no longer consistent with the donor from whom it was first established.

To know if a cell line is authentic or misidentified, it is consequently important to look at its genotype and not its behavior (phenotype). Indeed, the cell line’s phenotype can vary with the passage number and/or the culture conditions. Misidentification can be due to several causes.

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Whatever the problem, depending on the moment it occurs, a distinction is made between “false cell line” and “misidentified cell line”. If it happens early, during cell line establishment, no original material has been retained, and the cell line will be considered a false cell line. If it happens late, after the cell line has been established and distributed, the original material still exists. Only some stocks are therefore false and these will be defined as misidentified cells.

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Causes of misidentified cell lines‌

Consequences of working with the wrong cell line‌

What can you do to ensure working with the right cells?‌

What/ Who is ICLAC (International Cell Line Authentication Committee)?‌

Incorporating cell authentication testing into cell culture routine

Whereas no mandatory testing schedule exists at this time, it is recommended to systematically check and document cell line identity at least:
  • When a foreign cell line enters the cell culture laboratory.
  • During establishment of frozen cell stocks.
  • At the beginning and at the end of an experimental project.
  • Before distributing a cell line.

Choosing a cell authentication method

Several different methods are available to detect inter-species and intra-species cross-contamination in cell cultures. These methods verify cell line identity on the protein or genetic level. The acquisition and application of authentication methods is not difficult for trained scientists. However, reliable interpretation of the results requires advanced experience. Depending on the testing method, special laboratory equipment might also be necessary. For support, commercial services may be employed to perform testing and/or interpretation of analysis within a short turnaround time.

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Working with eukaryotic cells often requires the user to handle cell lines of different origins. Have you ever thought of cell identity and cell authentication? What has to be considered during cultivation as well as establishment of cell lines in the cell culture laboratory? Simply have a closer look at our Cell Handling Workflow to refresh your knowledge of the essential steps to ensure you are working with your cells of interest and that you avoid misidentification.

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References:
1. http://iclac.org/wp-content/uploads/ICLAC_Terms-of-Reference_2015_v2_6.pdf
2. Gartler (1967) Genetic markers as tracers in cell culture. Natl Cancer Inst Monogr. 26:167-95.
3. Gartler (1968) Apparent HeLa cell contamination of human heteroploid cell lines. Nature. 217:750-751.
4. Nelson-Rees and Flandermeyer (1977). Inter- and intraspecies contamination of human breast tumor cells HBC and BrCa5 and other cell cultures. Science. 195(4284):1343-4.
5. Lorsch et al. (2014) Cell Biology. Fixing problems with cell lines. Science. 346(6216):1452-3.
6. Marx (2014) Cell-line authentication demystified. Nature Methods. 11: 483-488.
7. http://www.celllineauthentication.com/journal-requirements.html
8. http://grants.nih.gov/grants/guide/notice-files/NOT-OD-15-103.html
9. Freedman et al. (2015) The culture of cell culture practices and authentication - Results from a 2015 survey. BioTechniques. 59:189-192.

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STR profiling‌

Karyotyping‌

Isoenzyme profiling‌

DNA barcoding‌