Characterized Cell Line Core Facility
About Cell Line Validation
Between 18 and 36% of cell lines are either misidentified or cross-contaminated1. In the NCI60 set, three lines are duplicates (MDAMB435/M14, MCF7-ADR/OVCAR8 and U373/SNB1.9). Genetic drift has introduced mutations in key regulatory pathways, leading to contradictory results from different laboratories. A recent notice from NIH requires cell line validation for grant applications to be considered of the highest quality2. Journals such as Science, Nature and PNAS are adopting requirements for cell line validation for publication. Cell lines that have been extensively characterized at the DNA, RNA and protein levels will allow investigators to choose the correct cell line for their research. Pre-characterized cell lines will decrease the cost to researchers since this will eliminate repeat analysis. Thus, cell line validation is a critical issue for both scientific publications and grant applications.
MD Anderson’s Cell Line Authentication Policy
MD Anderson now requires all researchers to validate their cell at least once per year. The information can be found in ACA#1044 which was sent by Dr. DuBois in December, 2010.
Short Tandem Repeat (STR) Method
The method that most journals require for validation of human cell lines is based on short tandem repeats (STR). STR repeats are regions of microsatellite instability with defined tri- or tetrad-nucleotide repeats that are located throughout the chromosomes. PCR reactions using primers on non-repetitive flanking regions will generate PCR products of different sizes based on the number of repeats in the region; the size of these PCR products are determined by capillary electrophoresis. By combining between 8 and 16 STR loci, such as D5S818, D13S317, D7S820, D16S539, vWA, TH01, TPOX, CSF1PO, it is possible to uniquely identify a sample. This is the same method that is used in forensics to match biological samples. We provide testing of STR on cell lines using the Promega Power Plex 16HS Kit.
If a human cell line has a known STR profile, the best and least expensive way to validate the cell line is to compare the STR profile against a database of known STR profiles. We have STR profiles from many different commercial sources, and we compare against this known commercial list.
If a human cell line does not have a known STR profile, we compare against all of the samples submitted to our core facility. All unique STR profiles are then added to our database for future MD Anderson reference. If there is a known mutation in the cell line, we may be able to confirm the mutation for an additional fee.
If the cell line is not a human cell line, we suggest that you get your cell line tested at the Molecular Cytogenetics core, 713-834-6395.
If the cell line is a potential mixture of a human cell line and a different species, we suggest that you get your cell line tested at the Molecular Cytogenetics core, 713-834-6395. The STR profiling will not detect any DNA other than human.
Mutation Detection (Sequenom)
Somatic mutations are responsible for transformation of normal cells into tumor cells. In addition to providing information on what is driving the cells to uncontrolled cell growth, the mutation pattern in a cell can help confirm the identity of a cell line. We use a Sequenom MALDI TOF MassArray system that can detect over 100 different common somatic mutations. As with all high throughput sequencing methods, we cannot guarantee these results. Please confirm all mutation calls with another technology. Also please note that in order to be certain that a base call is a somatic mutation, normal (non-tumor) DNA will also need to be screened.
We also offer mycoplasma detection using the MycoAlert kit from Lonza.
Purchase Cell Line
We have a limited number of cell lines that have been validated and tested for Mycoplasma contamination. Click here for the list of available cell lines.
Please contact Keri Sherman at CCSGCCLC@mdanderson.org or 713-792-5743.
Core Grant Citation
This facility is funded by NCI # CA016672. Publications should cite the Core grant in the acknowledgment section:
“STR DNA fingerprinting was done by the Cancer Center Support Grant-funded Characterized Cell Line core, NCI # CA016672.”
Two copies of the publication acknowledging the Core grant should also be submitted to the facility at Unit 950.
- Lacroix M. Persistent use of "false" cell lines. International Journal of Cancer, 2008. 122(1):1-4.
- Bravo NR, Gottesman M. Notice Regarding Authentication of Cultured Cell Lines. 2007 [cited 2008 1/28/08].
- Rae JM, et al. MDA-MB-435 cells are derived from M14 melanoma cells--a loss for breast cancer, but a boon for melanoma research. Breast Cancer Research & Treatment, 2007. 104(1):13-9.
- Chatterjee R. Cell biology. Cases of mistaken identity. Science, 2007. 315(5814):928-31.
- Drexler HG, et al. False leukemia-lymphoma cell lines: an update on over 500 cell lines. Leukemia, 2003. 17(2):416-26.
- MacLeod RA, et al. Widespread intraspecies cross-contamination of human tumor cell lines arising at source. International Journal of Cancer, 1999. 83(4):555-63.