SKY and genetic fingerprinting reveal a cross-contamination of the putative normal colon epithelial cell line NCOL-1

Melcher, Ralph; Maisch, Stefanie; Koehler, Sigrid; Bauer, Margit; Steinlein, Claus; Schmid, Michael; Kudlich, Theodor; Schauber, Jürgen; Luehrs, Hardi; Menzel, Thomas; Scheppach, Wolfgang

doi:10.1016/j.cancergencyto.2004.08.023

Cited by 13 publications

(6 citation statements)

References 13 publications

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“…This will permit easy verification of cell line identity to future investigators. Such verification has been strongly advocated to ensure the integrity of results with cell lines and xenografts [35], [36].…”

Section: Resultsmentioning

confidence: 99%

Characterization and Drug Resistance Patterns of Ewing's Sarcoma Family Tumor Cell Lines

et al. 2013

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Despite intensive treatment with chemotherapy, radiotherapy and surgery, over 70% of patients with metastatic Ewing's Sarcoma Family of Tumors (EFT) will die of their disease. We hypothesize that properly characterized laboratory models reflecting the drug resistance of clinical tumors will facilitate the application of new therapeutic agents to EFT. To determine resistance patterns, we studied newly established EFT cell lines derived from different points in therapy: two established at diagnosis (CHLA-9, CHLA-32), two after chemotherapy and progressive disease (CHLA-10, CHLA-25), and two at relapse after myeloablative therapy and autologous bone marrow transplantation (post-ABMT) (CHLA-258, COG-E-352). The new lines were compared to widely studied EFT lines TC-71, TC-32, SK-N-MC, and A-673. These lines were extensively characterized with regard to identity (short tandem repeat (STR) analysis), p53, p16/14 status, and EWS/ETS breakpoint and target gene expression profile. The DIMSCAN cytotoxicity assay was used to assess in vitro drug sensitivity to standard chemotherapy agents. No association was found between drug resistance and the expression of EWS/ETS regulated genes in the EFT cell lines. No consistent association was observed between drug sensitivity and p53 functionality or between drug sensitivity and p16/14 functionality across the cell lines. Exposure to chemotherapy prior to cell line initiation correlated with drug resistance of EFT cell lines in 5/8 tested agents at clinically achievable concentrations (CAC) or the lower tested concentration (LTC): (cyclophosphamide (as 4-HC) and doxorubicin at CAC, etoposide, irinotecan (as SN-38) and melphalan at LTC; P<0.1 for one agent, and P<0.05 for four agents. This panel of well-characterized drug-sensitive and drug-resistant cell lines will facilitate in vitro preclinical testing of new agents for EFT.

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Section: Resultsmentioning

confidence: 99%

Characterization and Drug Resistance Patterns of Ewing's Sarcoma Family Tumor Cell Lines

et al. 2013

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“…The researchers used spectral karyotyping to show this cell line is identical to LoVo, a cell line derived from a colon carcinoma (39). Yoshino et al used short tandem repeat (STR) polymorphism analysis to examine approximately 400 cell lines in the Cell Engineering Division of the Japanese research institution RIKEN and found that 10 of the human cell lines were identical to a different cell line that had been earlier deposited in the collection and that had been misidentified by the depositor (40).…”

Section: Cross-contamination and Misidentificationmentioning

confidence: 99%

The Costs of using Unauthenticated, Over-Passaged Cell Lines: How Much more Data do we Need?

Marshall²,

et al. 2007

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Increasing data demonstrate that cellular cross-contamination, misidentified cell lines, and the use of cultures at high-passage levels contribute to the generation of erroneous and misleading results as well as wasted research funds. Contamination of cell lines by other lines has been recognized and documented back to the 1950s. Based on submissions to major cell repositories in the last decade, it is estimated that between 18% and 36% of cell lines may be contaminated or misidentified. More recently, problems surrounding practices of over-subculturing cells are being identified. As a result of selective pressures and genetic drift, cell lines, when kept in culture too long, exhibit reduced or altered key functions and often no longer represent reliable models of their original source material. A review of papers showing significant experimental variances between low- and high-passage cell culture numbers, as well as contaminated lines, makes a strong case for using verified, tested cell lines at low- or defined passage numbers. In the absence of cell culture guidelines, mandates from the National Institutes of Health (NIH) and other funding agencies or journal requirements, it becomes the responsibility of the scientific community to perform due diligence to ensure the integrity of cell cultures used in research.

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“…Other methods such as HLA typing and spectral karyotyping may be used for further resolution (Melcher et al 2005).…”

Section: The Methods For Authenticationmentioning

confidence: 99%

“…Van Helden (1988) reported that the human esophageal squamous carcinoma cell lines HCu 10, 18, 22, 27, and 39 were genetically identical, while Ogura et al (1997) reported that lines JTC-3, OF, and OE isolated in 1959, 1969, and 1971, respectively, were HeLa cells. More recently, Melcher (2005) reported that the putative normal colon epithelial cell line NCOL-1 was probably derived from the colon carcinoma LoVo. Furthermore, spectral karyotyping and DNA fingerprinting revealed that a subline of NCOL 1 and LoVo were identical while another putative subline of NCOL 1 had additional markers.…”

Section: Brief Historymentioning

confidence: 98%

Eradication of cross-contaminated cell lines: A call for action

Nardone

2007

Cell Biol Toxicol

108

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This "white paper" was prepared and widely disseminated in an attempt to sound an alarm about the long-term existence of a grave, unresolved and growing problem that affects a significant portion of biomedical research, namely, the use of misidentified and cross-contaminated cell cultures. The "white paper" shows how bold action could bring about a profession-wide change in practice that would prevent further erosion. Misidentification and inter- and intra-specific cross-contamination of mammalian cell cultures used in research continues as a widespread problem despite an awareness that dates back more than 45 years. Awareness of the problem has led to a good understanding of the causes of cross-contamination and appropriate preventive measures. It has also led to the application of robust methods for the authentication of cell lines. Yet the problem continues unabated. Estimates of the incidence of research papers flawed by the use of misidentified and cross-contaminated cell cultures approximate 15-20%. The gravity of the situation calls for a strategy that would deliver a remedial message of authentication to virtually all cell culture researchers and also ensure compliance with the message. At the core of the strategy proposed herein is having cell line authentication as a condition for the award of research grants and for the publication of research findings.

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SKY and genetic fingerprinting reveal a cross-contamination of the putative normal colon epithelial cell line NCOL-1

Cited by 13 publications

References 13 publications

Characterization and Drug Resistance Patterns of Ewing's Sarcoma Family Tumor Cell Lines

Characterization and Drug Resistance Patterns of Ewing's Sarcoma Family Tumor Cell Lines

The Costs of using Unauthenticated, Over-Passaged Cell Lines: How Much more Data do we Need?

Eradication of cross-contaminated cell lines: A call for action

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