Cell Line Authentication – Why it Matters

Sub-optimal growth conditions, species misidentification, and the contamination of cultures with adventitious agents, are a few of the causes that can affect cell behavior and authenticity – all of which can lead to irreproducible results.
Cell line authentication should prove if your cell line is:
  • Comprised of the same cells as the original primary or continuous cell culture from which the cell line was derived
  • Free of cross-contaminating interspecies cells, and if possible, intraspecies cell types
  • Free of adventitious agents, such as mycoplasma, bacteria, fungi, viruses
  • Of expected morphology, ploidy, and phenotype under a given set of culture conditions
When should your cell lines be checked/authenticated?
  • On arrival at the facility, if not obtained from a reputable source/ repository with a certificate of authenticity
  • When establishing a cell bank or cell stock prepared at your facility
  • If marked phenotypic or morphological changes are observed during routine culture
  • All of the above should be done prior to publishing results based on the cell line (View Publication Requirements)
Available References:
  • Short tandem repeat profiling: part of an overall strategy for reducing the frequency of cell misidentification. Nims RW, Sykes G, Cottrill K, Ikonomi P, Elmore E. In Vitro Cell Dev Biol Anim 46:811-819, 2010. PMID: 20927602
  • ANSI/ATCC ASN-0002-2011 (2012) Authentication of human cell lines: Standardization of STR profiling. ANSI eStandards Store
  • Short tandem repeat profiling provides an international reference standard for human cell lines. Masters JR, Thomson 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. Proc Natl Acad Sci U S A. 2001 Jul 3;98(14):8012-7. PMID: 1416159
  • Recommendation of short tandem repeat profiling for authenticating human cell lines, stem cells, and tissues. Barallon R, Bauer SR, Butler J, Capes-Davis A, Dirks WG, Elmore E, Furtado M, Kline MC, Kohara A, Los GV, MacLeod RA, Masters JR, Nardone M, Nardone RM, Nims RW, Price PJ, Reid YA, Shewale J, Sykes G, Steuer AF, Storts DR, Thomson J, Taraporewala Z, AlstonRoberts C, Kerrigan L. In Vitro Cell Dev Biol Anim. 2010 Oct;46(9):727-32. doi: 10.1007/ s11626-010-9333-z PMID: 20614197
  • Authentication of Human Cell Lines by STR DNA Profiling Analysis. Reid Y, Storts D, Riss T, Minor L. In: Sittampalam GS, Coussens NP, Nelson H, Arkin M, Auld D, Austin C, Bejcek B, Glicksman M, Inglese J, Iversen PW, Li Z, McGee J, McManus O, Minor L, Napper A, Peltier JM, Riss T, Trask OJ Jr., Weidner J, editors. Assay Guidance Manual [Internet]. Bethesda (MD): Eli Lilly & Company and the National Center for Advancing Translational Sciences; 2004-. 2013 May 1. PMID: 23805434
  • Reproducibility: changing the policies and culture of cell line authentication. Freedman LP, Gibson MC, Ethier SP, Soule HR, Neve RM, Reid YA. Nat Methods. 2015 Jun;12(6):493-7. doi: 10.1038/nmeth.3403. No abstract available. PMID:26020501
  • The Science of cell culture – Maintaining phenotypic and genotypic heterogeneity. Goldsborough, Mindy. DDNews. 2016 Dec. http://www.ddn-news.com/news?newsarticle=11141
  • Characterization and authentication of cancer cell lines: an overview. Reid YA. Methods Mol Biol. 2011;731:35-43. doi: 10.1007/978-1-61779-080- 5_4. Review. PMID:21516396