Cell Line Validation
Between 18% and 36% of cell lines are either misidentified or cross-contaminated (list of misidentified cell lines). A recent notice from NIH requires cell line validation for grant applications to be considered of the highest quality. Journals such as Science, Nature and PNAS also require 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's policy (ACA#1044) now requires all researchers to validate their cell lines at least once per year (Our Core highly suggests to validate cell lines every 6 months).
*STR Fingerprinting Service
Short Tandem Repeat (STR) DNA profiling, also known as DNA fingerprinting, offers the greatest value for cell line authentication. The assay is based on screening regions of microsatellite instability with defined tri- or tetrad-nucleotide repeats located throughout the chromosomes. PCR reactions using primers on non-repetitive flanking those regions will generate PCR products of different sizes based on the number of repeats in the region; the size of these PCR products is determined by capillary electrophoresis. By combining between 8 and 16 STR loci, such as D5S818, D13S317, D7S820, D16S539, vWA, TH01, TPOX, and CSF1PO, it is possible to uniquely identify a sample. The CCLC assay screens 16 loci using the Promega Powerplex 16 HS kit. Our test also includes matching the STR profiles against an internal database comprised of public profiles and profiles that are unique to cell lines developed by MDACC investigators. Our current database has over 4000 profiles.
Mycoplasma Contamination Testing
Mycoplasma is a bacterial contaminant commonly found in cell culture. It is easy to spread from culture to culture, and unlike most other bacterial or fungal contaminants, mycoplasma contamination cannot be easily detected without further testing. Mycoplasma are bacteria that do not contain a traditional cell membrane and can alter the metabolic characteristics of cells in culture negatively affecting experimental results. We provide this mycoplasma determination service using the Lonza MycoAlert Kit.
Purchase Cell Lines
The CCLC has a List of Cell Lines that have been validated and tested for mycoplasma contamination that are available for purchase. Please see the Services page for pricing and instructions on requesting cell lines.
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 CCSG-funded Characterized Cell Line Core, NCI # CA016672.”
Two copies of the publication acknowledging the CCLC grant should also be submitted to the facility at Unit 1058.
Education & References
- 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.