STR DNA Typing of Human Cell Lines: Detection of Intra- and Interspecies Cross-Contamination

Part of the Methods in Molecular Biology book series (MIMB, volume 946)


Inter- and intraspecies cross-contaminations (CCs) of human and animal cells represent a chronic problem in cell cultures leading to false data. Microsatellite loci in the human genome harboring short tandem repeat (STR) DNA markers allow individualization of cell lines at the DNA level. Thus, fluorescence polymerase chain reaction amplification of STR loci D5S818, D13S317, D7S820, D16S539, vWA, TH01, TPOX, CSF1PO, and Amelogenin for gender determination is the gold standard for authentication of human cell lines and represents an international reference technique. The major cell banks of the USA, Germany, and Japan (ATCC, DSMZ, JCRB, and RIKEN, respectively) have built compatible STR databases to ensure the availability of STR reference profiles. Upon determination of an STR profile of a human cell line, the suspected identity can be proven by online verification of customer-made STR data sets on the homepage of the DSMZ institute. Furthermore, an additional tetraplex PCR has been established to detect mitochondrial DNA sequences of rodent cells within a human cell culture population. Since authentic cell lines are the main prerequisite for rational research and biotechnology, the next sections describe a rapid and reliable method available to students, technicians, and scientists for certifying identity and purity of human cell lines of interest.

Key words

Authentication Cross-contamination DNA STR typing Human cell lines mtDNA typing Misidentification Quality control 


  1. 1.
    Buehring GC, Eby EA, Eby MJ (2004) Cell line cross-contamination: how aware are mammalian cell culturists of the problem and how to monitor it? In Vitro Cell Dev Biol Anim 40:211–215PubMedCrossRefGoogle Scholar
  2. 2.
    MacLeod RAF, Dirks WG, Kaufmann M, Matsuo Y, Milch H, Drexler HG (1999) Widespread intra-species cross-contamination of human tumor cell line arising at source. Int J Cancer 83:555–563PubMedCrossRefGoogle Scholar
  3. 3.
    Liscovitch M, Ravid D (2007) A case study in misidentification of cancer cell lines: MCF-7/AdrR cells (re-designated NCI/ADR-RES) are derived from OVCAR-8 human ovarian carcinoma cells. Cancer Lett 245:350–352PubMedCrossRefGoogle Scholar
  4. 4.
    Nardone RM (2007) Eradication of cross-contaminated cell lines: a call for action. Cell Biol Toxicol 23:367–372PubMedCrossRefGoogle Scholar
  5. 5.
    Drexler HG, Dirks WG, Matsuo Y, MacLeod RAF (2003) False leukemia-lymphoma cell lines: an update on over 500 cell lines. Leukemia 17:416–426PubMedCrossRefGoogle Scholar
  6. 6.
    Schweppe RE, Klopper JP, Korch C, Pugazhenthi U, Benezra M, Knauf JA, Fagin JA, Marlow LA, Copland JA, Smallridge RC, Haugen BR (2008) Deoxyribonucleic acid profiling analysis of 40 human thyroid cancer cell lines reveals cross-contamination resulting in cell line redundancy and misidentification. J Clin Endocrinol Metab 93:4331–4341PubMedCrossRefGoogle Scholar
  7. 7.
    Masters JR, Thompson JA, Daly-Burns B, Reid YA, Dirks WG, Packer P, Toji LH, Ohno T, Tanabe H, Arlett CF, Kelland LR, Harrison M, Virmani A, Ward TH, Ayres KL, Debenham PG (2001) Short tandem repeat profiling provides an international reference standard for human cell lines. Proc Natl Acad Sci 98:8012–8017PubMedCrossRefGoogle Scholar
  8. 8.
    Sullivan KM, Mannucci A, Kimpton CP, Gill P (1993) A rapid and quantitative DNA sex test: fluorescence-based PCR analysis of X-Y homologous gene amelogenin. Biotechniques 15:636–641PubMedGoogle Scholar
  9. 9.
    White HW, Kusukawa N (1997) Agarose-based system for separation of short tandem repeat loci. Biotechniques 22:976–980PubMedGoogle Scholar
  10. 10.
    Dirks WG, MacLeod RAF, Nakamura Y, Kohara A, Reid Y, Milch H, Drexler HG, Mizusawa H (2010) Cell line cross-contamination initiative: an interactive reference database of STR profiles covering common cancer cell lines. Int J Cancer 126:302–304CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2013

Authors and Affiliations

  1. 1.Department of Human and Animal Cell LinesDSMZ, German Collection of Microorganisms and Cell Cultures and German Biological Resource CenterBraunschweigGermany

Personalised recommendations