Identification of Cell Lines
Cross contamination and errors in the identification of cell lines is estimated to be in the order of between 15% and 20%. This is a serious problem for correctly interpreting data from work carried out with these cells, as the work may not have been done with the supposed lines.
Working with properly identified materials prevents the loss of time and money, non-reproducible results, and the loss of credibility.
How are cell lines identified?
LThe guidelines and expert recommendations tell us that the best way of identifying cell lines is by performing an STR (short tandem repeat) allele profile and comparing it with well established databases. (Nature Reviews Cancer 10 (June 2010) | doi: 10.1038/nrc2852).
The STRs analyzed are amelogenin, CSF1PO, D13S317, D16S539, D5S818, D7S820, THO1, TPOX, vWA.
When should it be performed?
Cell-line identification is recommended in the following cases:
- If the phenotype of the cell line is not as described.
- Before starting a new series of experiments or publishing results.
- Before freezing for storage.
- When a new line is created.
- Every quarter, if the cell line is actively cultured.
A sample of cells is subjected to multiplex PCR to amplify the regions of the STRs. The amplifications are run through an ABI 3730xl Genetic Analyzer (ABI, St. Louis, MO) and the size of the fragments in each locus are identified using the GeneMapper v.4 (ABI) software package.
The profile of the STRs obtained for each cell line is compared with well-established databases in order to look for concordances with studied cell lines.