Multiplex human Cell Authentication (MCA)

Cell cultures are important tools for basic and applied biomedical research. About 15-20% of human cell lines have been estimated to be mislabelled or affected by cross-contamination with other human cell lines. The need for cell line authentication has been emphasized by granting agencies as well as by many high ranking journals (no cell authentication = no grant/publication).

Short tandem repeat (STR) profiling is widely used to verify the authentication of human cell lines. However, widely used cell lines (e.g. HEK293, CCRF-CEM, Jurkat, ...) with mutations in their mismatch repair genes have been shown to acquire changes in their STR profile upon long-term culture leading to false authentication results.

To solve these limitations, the DKFZ Core Facility "Contamination control" has developed in collaboration with the DSMZ (Braunschweig) a high-throughput Multiplex human Cell Authentication (MCA) test (Castro et al., submitted) and has built a SNP database for currently 547 distinct STR-authenticated human cell lines (overview of cells included in database). As little as 3% contaminating cells from another human cell line are detectable. MCA can also robustly authenticate widely used cell lines with mutations in their mismatch repair genes, such as hMutSα.

Non-DKFZ members have access to this service through the Steinbeis-Transfer Centre Multiplexion.

Multiplex Human Cell Line Authentication Service

Test specifications

Confirmation of human cell identity is done using specific primer sequences in a multiplex PCR targeting 24 SNP regions, and subsequent hybridisation using allele-specific oligonucleotide probes. The complete genotype information is compared to the database allowing to identify the correct cell line identity. Several positive and negative controls are included to test assay performance.

Application

• authentication of human cell lines
• detection of ongoing intra-human cross-contaminations
• authentication of newly-established human cell lines or sublines by genotyping paired donor and derived cell line samples

Procedure

• Purify cell line DNA (protocol)
• Adjust DNA concentration to 15 to 30ng/µL in SafeLock tubes (30µL required)
• Complete the Cell Authentication Submission Form (Login required)
• Label tubes with ‘SNP’ AND 'KstSt' AND 'specific sample name' as provided in the Submission Form
• Deliver DNA samples to the ATV building INF 242 every Monday, 10-12 a.m. to the doorman

Attention: The test will be performed in the week of the fourth Monday each month. No delivery of samples between December 27 to January 7. You should get your results within 4-5 working days. In December 2011 no testing will be performed.

• Multiplex PCR-based amplification of 24 SNP regions
• Luminex-based detection of genotypes by specific oligonucleotide probes
• Comparison of cell signature to database
• Transfer of results to customer (the report can be used as certificate for granting agencies and journals)

Recommended frequency of testing

According to our own experience, we recommend to control

• All batches of stocked cell lines once
• All new cell lines
• Long term-cultured cell lines every month
• Prior to or concomitant to storage in liquid nitrogen
• In case of alterations in cell features
• In case of problems with reproducibility of results

Please note: This DKFZ-internal service is designed to help you comply with safety regulations, to improve quality control and to help your research. It is NOT suited for clinical diagnostics.

For more information, please contact us.

For detailed cost information please consult the pricelist (login required).