Advertisement

Cytotechnology

, Volume 64, Issue 4, pp 373–378 | Cite as

The necessity of identity assessment of animal intestinal cell lines: A case report

  • Klaus G. SteubeEmail author
  • Anne-Leena Koelz
  • Cord C. Uphoff
  • Hans G. Drexler
  • Jeannette Kluess
  • Pablo Steinberg
Brief Report

Abstract

Eight intestinal cell lines, established from different animal species were submitted to DSMZ (German Collection of Microorganisms and Cell Cultures) in order to analyze their species of origin and their microbial contamination. Species identity was determined by PCR targeting mitochondrial genes and hence confirmed by sequencing the amplified PCR products. For three cell lines (CIEB, CLAB, PSI-1) we confirmed the species identity, whereas the species of origin of the three other cell lines (B6, B10XI and IPEC) was not the expected one: B6 and B10XI cells, which were supposed to be of chicken origin were identified as porcine cells. IPEC, allegedly a sub clone of the well-known porcine intestinal cell line IPEC-J2, was of bovine instead of porcine origin. However, two further IPEC-clones, namely IPEC-1 and IPEC-J2, provided by another source were shown to be derived from the correct species (i.e. pig). Furthermore, six out of these eight cell lines turned out to be highly contaminated with mycoplasma. Alerted by this high incidence of infected and false specified cell lines, we feel obliged to inform all those working with animal intestinal cell lines and we strongly recommend verifying the species identity before using them. Also, the presence of mycoplasma should be tested when taking the cells in culture for the first time, and this mycoplasma control should be repeated at regular time intervals (e.g. every 4 weeks).

Keywords

Cell Lines Identity control of cell lines Quality control of cell lines Species-PCR Speciation Mycoplasma Infection Intestinal cell lines IPEC-cells 

Notes

Acknowledgments

We gratefully thank Prof. Dr. Hermann-Josef Rothkötter, University of Magdeburg for donation of the IPEC-1 and IPEC-J2 cells to DSMZ.

References

  1. Allston-Roberts C, Barallon R, Bauer SR, Butler J, Capes-Davis A, Dirks WG, Elmore E, Furtado M, Kerrigan L, Kline MC, Kohara A, Los GV, MacLeod RAF, Masters JR, Nardone M, Nims RW, Price PJ, Reid YA, Shewale J, Steuer AF, Storts DR, Sykes G, Taraporewala Z, Thomson J (2010) Cell line misidentification: the beginning of the end. Nat Rev Cancer 10:441–448CrossRefGoogle Scholar
  2. Berschneider HM (1989) Development of normal cultured small intestinal epithelial cell lines which transport Na and Cl. Gastroenterol 96:A41Google Scholar
  3. Capes-Davis A, Theodosopoulos G, Atkin I, Drexler HG, Kohara A, MacLeod RAF, Masters JR, Nakamura Y, Reid YA, Reddel RR, Freshney RA (2010) Check your cultures! A list of cross-contaminated or misidentified cell lines. Int J Cancer 127:1–8CrossRefGoogle Scholar
  4. Chantret I, Barbat A, Dussaulx E, Brattain MG, Zweibaum A (1988) Epithelial polarity, villin expression, and enterocytic differentiation of cultured human colon carcinoma cells: a survey of twenty cell lines. Cancer Res 48:1936–1942Google Scholar
  5. Chatterjee R (2007) Cell biology: cases of mistaken. Science 315:928–931CrossRefGoogle Scholar
  6. Cooper JK, Sykes G, King S, Cottrill K, Ivanova NV, Hanner R, Ikonomi P (2007) Species identification in cell culture: a prolonged molecular approach. In Vitro Cell Dev Biol Anim 43:344–351CrossRefGoogle Scholar
  7. Diesing A-K, Nossol C, Panther P, Walk N, Post A, Kluess J, Kreutzmann P, Dänicke S, Rothkötter H-J, Kahlert S (2011) Mycotoxin deoxynivalenol (DON) mediates biphasic cellular response in intestinal porcine epithelial cell lines IPEC-1 and IPEC-J2. Toxicol Lett 200:8–18CrossRefGoogle Scholar
  8. Drexler HG, Dirks WG, Matsuo Y, MacLeod RAF (2003) False leukemia- lymphoma cell lines: an update on over 500 cell lines. Leukemia 17:416–426CrossRefGoogle Scholar
  9. Geens MM, Niewold TA (2011) Optimizing culture conditions of a porcine epithelial cell line IPEC-J2 through a histological and physiological characterization. Cytotechnolgy 63:415–423CrossRefGoogle Scholar
  10. Gonzalez-Vallina R, Wang H, Zhan R, Berschneider HM, Lee RM, Davidson NO, Black DD (1996) Lipoprotein and apolipoprotein secretion by a newborn piglet intestinal cell line (IPEC-1). Am J Physiol 271:G249–G259Google Scholar
  11. Lacroix M (2008) Persistent use of "false“ cell lines. Int J Cancer 122:1–4CrossRefGoogle Scholar
  12. Liu MY, Lin SC, Liu H, Candal F, Vafai A (2003) Identification and authentication of animal cell culture by polymerase chain reaction amplification and DNA sequencing. In Vitro Cell Dev Biol Anim 39:424–427CrossRefGoogle Scholar
  13. MacLeod RAF, Drexler HG (2006) Public repositories: users reluctant to give materials. Nature 439:912CrossRefGoogle Scholar
  14. MacLeod RAF, Dirks WG, Matsuo Y, Kaufmann M, Milch H, Drexler HG (1999) Widespread intraspecies cross-contamination of human tumor cell lines arising at source. Int J Cancer 83:555–563CrossRefGoogle Scholar
  15. MacLeod RAF, Dirks WG, Dexler HG (2002) Persistent use of misidentified cell lines and its prevention. Genes Chromosom Cancer 33:103–105CrossRefGoogle Scholar
  16. Mariani V, Palermo S, Fiorentini S, Lanubile A, Giuffra E (2009) Gene expression study of two widely used pig intestinal epithelial cell lines: IPEC-J2 and IPI-2I. Vet Immunol Immunopathol 131:278–284CrossRefGoogle Scholar
  17. Masters JR (2002) False cell lines: the problem and the solution. Cytotechnology 39:17–22CrossRefGoogle Scholar
  18. Nardonne RM (2007) Eradication of cross-contaminated cell lines. A call for action. Cell Biol Toxicol 23:367–372CrossRefGoogle Scholar
  19. Nelson-Rees WA, Daniels DW, Flandermeyer RR (1981) Cell cross-contamination in cell cultures. Science 212:446–452CrossRefGoogle Scholar
  20. Nossol C, Diesing AK, Walk N, Faber-Zuschratter H, Hertig R, Post A, Kluess J, Rothkötter HJ, Kahlert S (2011) Air-liquid interface cultures enhance the oxygen supply and trigger the structural and functional differentiation of intestinal porcine epithelial cells (IPEC). Histochem Cell Biol 36:103–115CrossRefGoogle Scholar
  21. Ono K, Motonobu S, Yoshida T, Ozawa Y, Kohara A, Takeuchi M, Mizusawa H, Sawada H (2007) Species identification of animal cells by nested PCR targeted to mitochondrial DNA. In Vitro Cell Dev Biol Anim 43:168–175CrossRefGoogle Scholar
  22. Pinton P, Braicu C, Nougayrede JP, Laffitte J, Taranu I, Oswald IP (2010) Deoxynivalenol impairs porcine intestinal barrier function and decreases the protein expression of claudin-4 through a mitogen-activated protein kinase-dependent mechanism. J Nutr 140:1956–1962CrossRefGoogle Scholar
  23. Schierack P, Nordhoff M, Pollmann M, Weyrauch KD, Amasheh S, Lodemann U, Jores J, Tachu B, Kleta S, Blikslager A, Tedin K, Wieler LH (2006) Characterization of a porcine intestinal epithelial cell line for in vitro studies of microbial pathogenesis in swine. Histochem Cell Biol 125:293–305CrossRefGoogle Scholar
  24. Stacey GN, Hoelzl H, Stephenson JR, Doyle A (1997) Authentication of animal cell cultures by direct visualization of repetitive DNA, aldolase gene PCR and isoenzyme analysis. Biologicals 25:75–85CrossRefGoogle Scholar
  25. Stacey GN, Masters JR, Hay RJ, Drexler HG, MacLeod RAF, Freshney RI (2000) Cell contamination leads to inaccurate data: we must take action now. Nature 403:356CrossRefGoogle Scholar
  26. Steube KG, Meyer C, Uphoff CC, Drexler HG (2003) A simple method using β-globin PCR for the species identification of animal cell lines. In Vitro Cell Dev Biol Anim 39:468–475CrossRefGoogle Scholar
  27. Steube KG, Koelz AL, Drexler HG (2008) Identification and verification of rodent cell lines by polymerase chain reaction. Cytotechnology 56:49–65CrossRefGoogle Scholar
  28. Stevenson R (1987) Development of cell banking in the US 1960–1985: a strategic approach to quality control. Adv Cell Cult 51:267–288Google Scholar
  29. Uphoff CC, Drexler HG (2011a) Detecting Mycoplasma contamination in cell cultures by polymerase chain reaction. In: Cree I (ed) Cancer cell culture: methods and protocols, 2nd edn. Springer, Berlin, pp 93–103CrossRefGoogle Scholar
  30. Uphoff CC, Drexler HG (2011b) Elimination of mycoplasmas from infected cell lines using antibiotics. In: Cree I (ed) Cancer cell culture: methods and protocols, 2nd edn. Springer, Berlin, pp 105–114CrossRefGoogle Scholar
  31. Zoon KC (1993) Points to consider in the characterization of cell lines used to produce biologicals. Center for Biological Evaluation and Research. Food and Drug Administration, Rockville, MD, pp 7–8Google Scholar

Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Klaus G. Steube
    • 1
    Email author
  • Anne-Leena Koelz
    • 1
  • Cord C. Uphoff
    • 1
  • Hans G. Drexler
    • 1
  • Jeannette Kluess
    • 2
  • Pablo Steinberg
    • 3
  1. 1.Leibniz-Institut, DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbHBraunschweigGermany
  2. 2.Medical Faculty, Institute of AnatomyOtto-von-Guericke UniversityMagdeburgGermany
  3. 3.Institute for Food Toxicology and Analytical ChemistryUniversity of Veterinary Medicine HannoverHannoverGermany

Personalised recommendations