Literatur-Übersicht Kreuzkontaminationen in der Zellkultur

Kreuzkontaminationen sind neben den Mycoplasmen vielleicht das schlimmste Problem das die Zellkultur hat. Warum ? Weil es einen Grund hat, dass man mit einer bestimmten Zelllinie arbeitet. Wenn sich nun aber nach Jahren herausstellt, dass es gar nicht die gewollte Zelllinie ist ? Wenn man statt mit Brustkrebszellen mit Melanomzellen arbeitet ? Der Schaden dieser Kreuz-Kontamination für die Arbeitsgruppen, aber vor allem auch für die wissenschaftlichen Literatursammlungen ist verheerend! Tausende von Artikeln mit den falschen Zelllinien ... kein Wunder, dass man manches nicht reproduzieren kann!

Die aktuelle Liste der zur Zeit als kreuzkontaminiert nachgewiesenen Zellen von A. Capes Davis und I.R. Freshney kann man hier herunterladen.

Zellkultur Kreuz-Kontamination Entstehung

Zelllinien Authentifizierungs-Aufrufe

Besonders in den letzten Jahren häufen sich die Aufrufe namhafter Wissenschaftler und vor allem den Mitarbeitern von internationalen Zellbanken, dass nun endlich nach fast 50 Jahren Nutzung von kreuzkontaminierten Zelllinien eine Authentifizierung von Linien in allen Labors durchgeführt werden soll.

Check your cultures! A list of cross-contaminated or misidentified cell lines.

Capes-Davis A, Theodosopoulos G, Atkin I, Drexler HG, Kohara A, MacLeod RA, Masters JR, Nakamura Y, Reid YA, Reddel RR, Freshney RI.: Int J Cancer. 2010 Jul 1;127(1):1-8. Review

currently the list consists of 360 cross-contaminated cell lines
old study by DSMZ cited: 18% of 252 submitted cell lines were cross-contaminated
newer study by DSMZ cited: of 598 lymphoma cell lines previously submitted: authentic/myco− (n = 411, 69%); authentic/myco+ (n = 108, 18%); false/myco− (n = 41, 7%) and false/myco+ (n = 38, 6%)

Human leukemia and lymphoma cell lines as models and resources.

MacLeod RA, Nagel S, Scherr M, Schneider B, Dirks WG, Uphoff CC, Quentmeier H, Drexler HG.: Curr Med Chem 2008;15(4):339-59. Review

Characterization and authentication of cancer cell lines: an overview.

Reid YA.: Methods Mol Biol 2011;731:35-43. Review.

Konsequenzen der Nutzung nicht-authentifizierter Zellen

Die Konsequenzen der Nutzung von falsch-identifizierten Zelllinien ist unüberschaubar hoch. Für einzelne Arbeitsgruppen oder Personen bedeutet das "nur" den Verlust der Validität ihrer Aussagen, für die Wissenschaft im Allgemeinen jedoch, das die Literatur übersäht ist, mit falschen Daten.

Persistent use of "false" cell lines.

Lacroix M.: Int J Cancer 2008 Jan 1;122(1):1-4. Review

The costs of using unauthenticated, over-passaged cell lines: how much more data do we need?

Hughes P, Marshall D, Reid Y, Parkes H, Gelber C.: Biotechniques 2007 Nov;43(5):575, 577-8, 581-2 passim. Review

Aus diesem Artikel haben wir folgende Tabelle übernommen, modifiziert und ergänzt:

Year	Cell Cultures Observed	Results	Reference
1967	20 human cell lines tested.	18 were contaminated with HeLa.	Gartler SM. 1967. Natl Cancer Inst. Monogr. 16:1967.
1968	18 independent human cell lines.	18 lines were contaminated with HeLa.	Melcher, R., S. Maisch, S. Koehler, M. Bauer, C. Steinlein, M. Schmid, T. Kudlich, J. Schauber, et al. 2005. SKY and genetic fingerprinting reveal a cross-contamination of the putative normal colon epithelial cell line NCOL-1. Cancer Genet. Cytogenet. 151:84-87.
1973	Two cell lines claimed to be of human origin.	Both shown to be of mouse origin.	Stulberg, C.S. 1973. Extrinsic cell contamination of tissue cultures, p. 1-27. In J. Fogh (Eds.), Contamination in Tissue Culture. Academic Press, New York.
1974	Twenty independent cell lines.	Nine of the 20 cell cultures displayed HeLa markers.	Nelson-Rees, W.A., R.R. Flandermeyer, and P.K. Hawthorne. 1974. Banded marker chromosomes as indicators of intraspecies cellular contamination. Science 184:1093-1096.
1974	Two purported human carcinoma cancer lines (HTB-3 and HTB-39B) and 1 HEK cell line.	All three lines were HeLa cells.	Culliton, B.J. 1974. HeLa cells: contamination cultures around the world. Science 184:1058-1059.
1976	466 cell lines tested.	9% were shown to be cross-contaminated.	Nelson-Rees W. 1976. Science 191:1976.
1976	246 lines investigated specifically for evidence of cellular cross-contamination or mislabeling.	Overall 30% of the lines were incorrectly designated (14% for interspecies and 25% for intraspecies contamination).	Stulberg, C.S., W.D. Peterson, and W.F. Simpson. 1976. Identification of cells in culture. Am. J. Hematol. 1:237-242.
1977	279 cell lines from 45 laboratories.	In total, 41 lines not as purported; 21 were of the wrong species, 15 were HeLa instead of other human cell lines, one rodent cell line was contaminated with manta cells, one purported normal diploid human cell line was actually a BT-20 breast carcinoma cell line, and three purported mixed species lines lacked one of two species (one lacked rat in an avian-rat mixture; two lacked mouse in a human-mouse mixture).	Nelson-Rees, W.A. and R.R. Flandermeyer. 1977. Inter- and intraspecies contamination of human breast tumor cell lines HBC and BrCa5 and other cell cultures. Science 195:1343-1344.
1979	Two established human breast carcinoma cell lines of metastatic origin.	Both shown to be cross-contaminated.	Siciliano, M.J., P.E. Barker, and R. Cailleau. 1979. Mutually exclusive genetic signatures of human breast tumor cell lines with a common chromosomal marker. Cancer Res. 39:919-922.
1981	Four Hodgkin’s disease human cell lines (FQ, RB, SpR, and Ry).	None of the cell lines were Hodgkin’s. Three of the cell lines were shown to be identical. The origin of the fourth (RY) could not be established with certainty.	Harris, N.L., D.L. Gang, S.C. Quay, S. Poppema, P.C. Zamecnik, W.A. Nelson-Rees, and S.J. O’Brien. 1981. Contamination of Hodgkin’s disease cell cultures. Nature 289:228-230.
1981	From 103 sources, lines derived from endometrium, amniotic cells, breast carcinoma, and other gynecologic cancers, larynx, and lung cancer, gastrointestinal cancer, liver and bone-marrow-derived leukemia and lymphoma lines, urologic cancer, etc.	About 100 documented events of contamination.	Nelson-Rees, W.A., D.W. Daniels, and R.R. Flandermeyer. 1981. Cross-contamination of cells in culture. Science 212:446-452.
1984	257 cultures.	Overall 35% were contaminated; 36% of the human lines were cross-contaminated (25% by cells of another species and 11% by another human cell line).	Hukku, B., D. Halton, and M. Mally. 1984. Cell characterization by use of multiple genetic markers, p. 13-31. In R.T. Acton and D.J. Lynn (Eds.), Eukaryotic Cell Cultures. Plenum Press, New York.
1988	Insulin-producing cell line, clone 16, thought to be derived from human fetal endocrine pancreatic cell.	Documented to be of Syrian hamster origin.	Matsuba, I., A. Lernmark, and O. Madsen, B. Michelsen, J.H. Nielsen, J. Scholler, H. Vissing, B. Welinder, et al. 1988. Gene probes to detect cross-culture contamination in hormone producing cell lines. In Vitro Cell. Dev. Biol. 24:1971-1976.
1993	Sister cell lines SPI-801 and SPI-802 thought to be established from a patient with acute lymphoblastic leukemia (ALL).	Shown to be subclones of K-562, a chronic myeloid leukemia cell line.	Gignac, S.M., K. Steube, and L. Schleithoff. 1993. Multiparameter approach in the identification of cross-contaminated leukemia cell lines. Leuk. Lymphoma 10:359-368.
1994	Macrophage-monocyte U-937.	Found to be contaminated with K-562.	Reid, Y.A., K. O’Neill, and T.R. Chen. 1995. Cell cross-contamination of U-937. J. Leukoc. Biol. 57:804.
1999	189 cell cultures received by the German Collection of Microorganisms and Cell Cultures (DSMZ) cell bank (1990–1999) representing 170 human hematopoietic lines.	Seventeen out of 117 (14.5%) from original source and 11 out of 72 (15.3%) from a secondary source were shown to be cross-contaminated with another hematopoietic cell line.	Drexler, H.G., W.G. Dirks, and R.A.F. MacLeod. 1999. False human hematopoietic cell lines: cross-contaminations and misinterpretations. Leukemia 13:1601-1607.
1999	252 human cell lines submitted to the German DSMZ cell bank.	45 of the cell lines were contaminated, mostly by intraspecies contamination.	MacLeod, R.A.F., W.G. Dirks, Y. Matsuo, M. Kaufmann, H. Milch, and H.G. Drexler. 1999. Widespread intraspecies cross-contamination of human tumor cell lines arising at source. Int. J. Cancer 83:555-563.
2000	Cell line, ECV 304, a putative human endothelial line of umbilical vein origin.	Shown to be identical to cell line, T24/83 derived from human urinary bladder carcinoma.	Brown, J., S.J. Reading, S. Jones, C.J. Fitchett, J. Howl, A. Martin, C.L. Longland, F. Michelangeli, et al. 2000. Critical evaluation of ECV304 as a human endothelial cell model defined by genetic analysis and functional responses: a comparison with the human bladder cancer derived epithelial cell line T24/83. Lab. Invest. 80:37-45.
2002	TI-1 cell line reportedly established from peripheral blood blasts from male patient.	Shown to be a cross-contaminant of K-562, a line derived from bone marrow of a 53-year-old female patient with chronic myelogenous leukemia (CML).	Rush, L.J., K. Heinonen, K. Mrózek, B.J. Wolf, M. Abdel-Rahman, J. Szymanska, P. Peltomäki, F. Kapadia, et al. 2002. Comprehensive cytogenetic and molecular genetic characterization of TI-1 acute myeloid leukemia cell line reveals cross-contamination with K-562 cell line. Blood 99:1874-1876.
2002	440 leukemia cell lines tested for mycoplasmas and cross-contamination.	Many lines are misidentified and mycoplasma-contminated.	H. G. Drexler, C. C. Uphoff, W. G. Dirks, and R. A. F. MacLeod, “Mix-ups and mycoplasma: the enemies within,” Leukemia Research, vol. 26, no. 4, pp. 329–333, 2002.
2003	550 human leukemia-lymphoma cell lines.	Unequivocal evidence of misidentification for 82 (14.9%) of the lines was found.	Drexler, H.G., W.G. Dirks, Y. Matsuo, and R.A.F. MacLeod. 2003. False leukemia-lymphoma cell lines: an update on over 500 cell lines. Leukemia 17:416-426.
2005	Normal colon epithelial cell line NCOL-1.	Identified as the colon carcinoma cell line LoVo.	Gartler, S.M. 1968. Apparent HeLa cell contamination of human heteroploid cell lines. Nature 217:750-751.
2006	Approximately 400 human cell lines deposited in the RIKEN BioResource Center.	Ten human lines were identical to a different cell line in the collection.	Yoshino, K., E. Iimura, K. Saijo, S. Iwase, K. Fukami, T. Ohno, Y. Obata, and Y. Nakamura. 2006. Essential role of gene profiling analysis in the authentication of human cell lines. Hum. Cell 19:43-48.
2007	Human breast adenocarcinoma cell line MCF-7 and the derived line MCF7/AdrR.	Both were found to have originated from the ovarian adenocarcinoma cell line OVCAR-8.	Liscovitch, M. and D. Ravid. 2007. A case study in misidentification of cancer cell lines: MCF/AdrR cells (re-designated NCI/ADR-RES) are derived from OVCAR-8 human ovarian carcinoma cells. Cancer Lett. 245:350-352.
2007	Comparision of MDA-MB-435 with melanoma M14 cells.	Ongoing discussion. MDA-MB-435 are of M14 origin ? Compare to 2009 article.	MDA-MB-435 cells are derived from M14 melanoma cells-a loss for breast cancer, but a boon for melanoma research. Rae JM, Creighton CJ, Meck JM, Haddad BR, Johnson MD. Breast Cancer Res Treat. 2007 Jul;104(1):13-9.
2007	100 cell lines deposited at the National Cell Bank of Iran.	Eighteen of the lines were cross-contaminated.	Azari, S., N. Ahmadi, M.J. Tehrani, and F. Shokri. 2007. Profiling and authentication of human cell lines using short tandem repeat (STR) loci: report from the national cell bank of Iran. Biologicals 35:195-202.
2009	Literature review. Comparision of MDA-MB-435 with melanoma M14 cells.	MDA-MB435 and M14 are identical but both seem to be MDA-MB-435 due to female gender.	MDA-MB-435 and M14 cell lines: identical but not M14 melanoma? Chambers AF. Cancer Res. 2009 Jul 1;69(13):5292-3.