Cross-contamination and misidentification of cell lines is estimated to be in the range of 15-20%. This is a serious problem for the correct experiment interpretation, since these cell lines used for research are not what they are supposed to be.
Secugen offers now the possibility of testing the identity of the cellular lines you work with. Working with well identified materials avoid waste of time and money, non-reproducible results and loss of credibility.

How is the identification done?

The experts guidelines and recommendations say that the best way to identify cell lines is making a STR (Short Tandem Repeats) allele profile and compare it to well established databases. (Nature Reviews Cancer 10, (June 2010) | doi:10.1038/nrc2852). The STR analyzed in Secugen are Amelogenin, CSF1PO, D13S317, D16S539, D5S818, D7S820, THO1, TPOX, v WA.

When do it?

Cell line identification is recommended to be done:

• If the phenotype of the cell line is not the expected.
• Before starting a new serie of experiments.
• Before freezing to store.
• At early passage of new cell line establishment in order to characterize it.
• Quarterly if the cell line is grown actively.

Method of Analysis

On the sample of cells sent to Secugen, multiplex PCR is performed to amplify the STRs regions. The amplicons generated are run in an ABI 3730xl Genetic Analyzer (ABI, St. Louis, MO). The sizing of the STR fragments is performed with the software GeneMapper v.4 (ABI).
The STR profile obtained for each cell line is compared against well-established databases to find concordances with characterized cell lines.