Cell Line STR Profiling - Human Cell Line Authentication Services
Cell lines are often used in biomedical research to investigate disease pathways and therapies.
In late 2007, an open letter by a group of scientists led by Dr. Roland Nardone spurred the National Institutes of Health (NIH) to recommend authenticating cell lines in scientific studies.
In early 2012, the ATCC issued its standard to address the problem of cell line misidentification and "delineate a standardized, universally applicable method for authenticating new and established human cell lines and human primary tissue used in research."
The International Cell Line Authentication Committee (ICLAC) recommends that cell line STR profiling in accordance with the standard ASN-0002-2011. Recommendations from the standard should be followed, including the use of at least eight core STR loci and application of match criteria (80% match threshold). This threshold allows for possible drift in some cell lines.
DDC Medical recently increased their core testing capabilities by examining 22 polymorphic loci for human and cell line STR profiling. DDC Medical continues to lead the industry by being one of the first to unilaterally introduce this upgraded capability, at no additional cost to the researcher.
Studies have shown an alarming percentage of human cell lines, those purchased from commercial repositories and primary cell lines, are contaminated or misidentified. Poor cell culture technique, shared equipment and poor quality control in a laboratory can all contribute to the misidentification and contamination issues. In addition, genetic instability of some cell lines can result in changes at the DNA level that can compromise research results. Articles published on these topics indicate as much as 20 to 33 percent of all cell lines are misidentified or contaminated, but there is a simple and affordable solution.
DDC Medical, a division of DNA Diagnostics Center (DDC), one of the world’s largest DNA laboratories, offers Human Cell Line Authentication and Mycoplasma Detection services.
Peer review Journals throughout the research community recommends Cell Line Authentication and Mycoplasma Detection as part of the publication process. Grant providers are also aware of the issue and requiring Cell Line Authentication and Mycoplasma Detection ensures a sound investment.
DDC Medical Customers
• Academia Researchers and Teaching Labs
• Pharma Labs and Researchers
• Biotech Labs and Researchers
• Government Labs and Researchers
• Other University, private and non-profit cancer and cell-based research institutions
When to Conduct Cell Line Authentication
• When a new line is first established or purchased
• As a control measure at the start of a new study and concluding a study
• At least once every 6 months in active culture
• Before freezing down for storage
• When an unexpected result occurs
About the Test
Genetic instability of some cell lines can result in changes at the DNA level that can compromise research results. Cell Line Authentication will ensure the research is being conducted on the correct cells, maximizing reliable results, supporting publication efforts, and reducing the risk of wasted time and resources. DDC Medical provides testing and certification that many Journals, research funding agencies and the NIH strongly recommend or require.
Contamination can occur at any time so it is important to test frequently. As referenced by the ATCC's Cell Line Authentication standards (ASN-0002), the study "Short tandem repeat profiling: part of an overall strategy for reducing the frequency of cell misidentification"1 recommends cell line authentication:
"when a cell line is received from an outside source (repository, other investigator). An aliquot could be tested at the time of preparation of the initial frozen cell stock. The culture should have its profile reconfirmed after expanding two or three passages, to check if any contaminants previously below the detection threshold have grown to be more evident. Cells from the initial frozen stock may then be used for experiments with confidence that the identity is correctly established. The identity of the cell line should be reconfirmed when new cell stocks are prepared. For newly established cultures, a sample of the culture at the time of the first cryopreservation should be maintained to establish an authentication reference for later passages of the culture. If many different cell lines are employed within a given laboratory, check the identity of the cultures periodically, as such conditions increase the cross-contamination risk. The identity of cell lines used for studies to be published should be confirmed as part of the manuscript submission process".1
1 Source: Nims, R. W., Sykes, G., Cottrill, K., Ikonomi, P., and Elmore, E. 2010. Short tandem repeat profiling: part of an overall strategy for reducing the frequency of cell misidentification.
DDC Medical's Human Cell Line Authentication service examines 22 highly polymorphic genetic markers (loci) using STR DNA Analysis, exceeding the recommended 8 by the ATCC. In addition, each test performed by DDC Medical uses the DUAL Process™ which ensures every test results is accurate. A DNA profile is generated which uniquely identifies the cell line used in the research.
Each locus is characterized by a motif of repeat units typically 4 bases in length. The number of repeats detected in a DNA fragment identifies each allele. For example, if the designation at the D5S818 locus is 14, 16, two DNA fragments were detected; one with 14 repeats and one with 16 repeats. The loci used for Cell Line Authentication are utilized for human identification for paternity and forensic analysis and are not designed to detect non-human DNA. Cell lines established from different human sources will possess different DNA profiles.
An electropherogram provides the data generated by the STR DNA alalysis. This data shows the alleles detected at each examined location and allows you to see the plot of results via electrophoresis automatic sequencing.
This additional service compares the cell line to the ATCC's STR Profile database to determine if there is a known match (or misidentification) and provide the % match at ≥80% (per the ATCC ASN-0002 guidelines) - we do not report cell line matches below a 55% to 79% match as there is considerable doubt of relatedness.
- Electropherogram - An electropherogram provides the data generated by the STR DNA analysis. This data shows the alleles detected at each examined location and allows you to see the plot of results via electrophoresis automatic sequencing.
- Comparative Analysis
DDC Medical staff will enter the designated loci (plus amelogenin) from the STR profile generated into the available ATCC or DSMZ STR Database. A report will be generated indicating a match criterion based on an algorithm that compares your profile to the database. Cell lines can be an identical match or a match > 80% which would indicate that the cell lines are "related".
These services can be purchased with any DDC Medical Cell Line Authentication test. This analysis is important if; 1) you need to confirm the cells you are working with at the beginning of research your research 2) you need to validate the cell line before submitting an application to publish your findings.
DDC Medical offers competitive industry pricing, volume discounts and unique customized solutions to fit any need. Please complete the Estimate Request and a response will be sent within 24 hours. If you have an immediate need or have further questions, please contact DDC Medical at firstname.lastname@example.org or call 1-888-362-3928.