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Retracted: Marker-independent identification of glioma-initiating cells

A Retraction to this article was published on 27 September 2013

This article has been updated

Abstract

Tumor-initiating cells with stem cell properties are believed to sustain the growth of gliomas, but proposed markers such as CD133 cannot be used to identify these cells with sufficient specificity. We report an alternative isolation method purely based on phenotypic qualities of glioma-initiating cells (GICs), avoiding the use of molecular markers. We exploited intrinsic autofluorescence properties and a distinctive morphology to isolate a subpopulation of cells (FL1+) from human glioma or glioma cultures. FL1+ cells are capable of self-renewal in vitro, tumorigenesis in vivo and preferentially express stem cell genes. The FL1+ phenotype did not correlate with the expression of proposed GIC markers. Our data propose an alternative approach to investigate tumor-initiating potential in gliomas and to advance the development of new therapies and diagnostics.

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Figure 1: Glioma tumors and cell cultures contain an autofluorescent population.
Figure 2: FL1+ cells display exclusive self-renewal abilities.
Figure 3: Expression of stemness related genes in FL1+ cells.
Figure 4: FL1+ cells initiate and sustain tumor growth in vivo.
Figure 5: Tumorigenic capacity of FL1+ cells.

Change history

  • 08 September 2013

    In the version of this article initially published, we described a method to identify tumor-initiating cells from human primary gliomasphere cell cultures or directly from fresh human glioma specimens. We used FACS analysis to describe a tumor-initiating cell subpopulation that displayed a specific morphology (high forward scatter, low side scatter) and had high fluorescence in the FL1 channel of the FACS (excitation at 488 nm, emission at 520 nm). We have since become aware that the majority of the primary gliomasphere lines (7 of 10) used in this paper were contaminated with HEK cells expressing GFP. We had used these sphere cultures to illustrate the selection method, to measure self-renewal, to assess the expression of stemness genes and to test for tumorigenicity. As assessed by microsatellite analysis of 15 short tandem repeats, seven of the primary sphere cultures used in this paper and all of the three lines used for tumorigenicity experiments did not match their parental tissue, and their genetic profile was consistent with that of HEK cells. We further observed expression of GFP mRNA with reverse-transcription PCR in 7 of the 10 gliomasphere lines. Two of these lines (GSM-1 and OA-III-1) were traceable by microsatellite analysis to their parental tissue at low passages (2–9) but not at higher passages (>20), a result showing that contamination occurred during passage of the cells in the laboratory. We also described experiments done with cells prospectively isolated from freshly resected glioma tissues; we believe that these experiments remain valid. We discussed in the paper that these cells were rare and did not have the same high fluorescence as gliomasphere cultures. We speculated that glioma cells acquire high fluorescence under in vitro conditions. This led to our misinterpretation of the progressive increase in fluorescence of two of the gliomasphere cultures that were in fact being progressively overgrown by contaminating GFP-expressing HEK cells. All together, when we exclude HEK-contaminated cultures, we must conclude that glioma-initiating cells do not have high autofluorescence levels or acquire them during culture. Selection of a tumorigenic fraction may be possible on the basis of morphological characteristics. However, because an important part of the study was done on contaminated cells, we wish to retract the paper. We deeply apologize to the scientific community for erroneously reporting an artifactual phenomenon as well as for the delay in detecting, characterizing and reporting the error. Cross-contamination of cell cultures is likely to be a frequent problem. Routine tracing of cell lines by microsatellite analysis has been advocated before publishing work with cell lines. Our experience shows that this advice holds true also for primary cells passaged in culture.

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Acknowledgements

This work was supported by the foundations “Anita et Werner Damm-Etienne” and “Ernest and Lucie Schmidheiny” and in part by the Centre d'Imagerie Biomedicale of the Ecole Polytechnique Fédérale de Lausanne, the University of Lausanne, the University of Geneva and by the Leenaards and Jeantet foundations, as well as by Marie Curie action of the European Union (MRTN-CT-2006-035801) and the Swiss National Science Foundation (3100AO-108266/1). We thank K. Burkhardt for providing access to tissue sample collection and S.G. Clarkson for critical input.

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Authors and Affiliations

Authors

Contributions

I.R. and V.C. conceived the study and designed the experiments. V.C., D.M., M.-F.H., C.C. and V.M. performed all the experiments. I.R. and V.C. wrote the manuscript. P.-Y.D., M.E.H., R.G., N.d.T., V.C. and I.R. discussed, revised and checked the manuscript.

Corresponding author

Correspondence to Ivan Radovanovic.

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Competing interests

V.C. and I.R. are authors of a patent (PCT/IB2008/054872) related to the technology described in this article and filed by the University of Geneva and Geneva University Hospitals. V.C., I.R. and D.M. are founders and shareholders of Stemergie Biotechnology, Inc.

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Supplementary Figures 1–10 and Supplementary Tables 1–11 (PDF 2544 kb)

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Clément, V., Marino, D., Cudalbu, C. et al. Retracted: Marker-independent identification of glioma-initiating cells. Nat Methods 7, 224–228 (2010). https://doi.org/10.1038/nmeth.1430

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