Skip to main content

Advertisement

Log in

Mycoplasma Infection Alters Cancer Stem Cell Properties in Vitro

  • Published:
Stem Cell Reviews and Reports Aims and scope Submit manuscript

Abstract

Cancer cell lines can be useful to model cancer stem cells. Infection with Mycoplasma species is an insidious problem in mammalian cell culture. While investigating stem-like properties in early passage melanoma cell lines, we noted poorly reproducible results from an aliquot of a cell line that was later found to be infected with Mycoplasma hyorhinis. Deliberate infection of other early passage melanoma cell lines aliquots induced variable and unpredictable effects on expression of putative cancer stem cell markers, clonogenicity, proliferation and global gene expression. Cell lines established in stem cell media (SCM) were equally susceptible. Mycoplasma status is rarely reported in publications using cultured cells to study the cancer stem cell hypothesis. Our work highlights the importance of surveillance for Mycoplasma infection while using any cultured cells to interrogate tumor heterogeneity.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Abbreviations

CSC:

Cancer stem cell

SCM:

Stem cell media

References

  • Aggelis, V., Craven, R. A., Peng, J., Harnden, P., Cairns, D. A., Maher, E. R., & Banks, R. E. (2009). Proteomic identification of differentially expressed plasma membrane proteins in renal cell carcinoma by stable isotope labelling of a von hippel-lindau transfectant cell line model. Proteomics, 9(8), 2118–2130.

    Article  CAS  PubMed  Google Scholar 

  • Ailles, L. E., & Weissman, I. L. (2007). Cancer stem cells in solid tumors. Current Opinion in Biotechnology, 18(5), 460–466.

    Article  CAS  PubMed  Google Scholar 

  • Ali, N., Allam, H., May, R., Sureban, S. M., Bronze, M. S., Bader, T., & Houchen, C. W. (2011). Hepatitis C virus-induced cancer stem cell-like signatures in cell culture and murine tumor xenografts. Journal of Virology, 85(23), 12292–12303. doi:10.1128/jvi.05920-11.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Anaka, M., Freyer, C., Gedye, C., Caballero, O., Davis, I. D., Behren, A., & Cebon, J. (2012). Stem cell media culture of melanoma results in the induction of a nonrepresentative neural expression profile. [research support, Non-U.S. Gov’t]. Stem Cells, 30(2), 336–343. doi:10.1002/stem.786.

    Article  CAS  PubMed  Google Scholar 

  • Callaway, E. (2014). Contamination hits cell work. [News]. Nature, 511(7511), 518. doi:10.1038/511518a.

    Article  CAS  PubMed  Google Scholar 

  • Dennis Jr., G., Sherman, B. T., Hosack, D. A., Yang, J., Gao, W., Lane, H. C., & Lempicki, R. A. (2003). DAVID: database for annotation, visualization, and integrated discovery. Genome Biology, 4(5), P3.

    Article  PubMed  Google Scholar 

  • Drexler, H., & Uphoff, C. (2000). Contamination of cell culture, mycoplasma. In E. Spier (Ed.), Encyclopedia of cell technology (pp. 609–627). New York: Wiley.

    Google Scholar 

  • Gedye, C., Quirk, J., Browning, J., Svobodova, S., John, T., Sluka, P., & Davis, I. D. (2009). Cancer/testis antigens can be immunological targets in clonogenic CD133+ melanoma cells. [research support, Non-U.S. Gov’t]. Cancer Immunology, Immunotherapy, 58(10), 1635–1646. doi:10.1007/s00262-009-0672-0.

    Article  CAS  PubMed  Google Scholar 

  • Gibbs, P., Hutchins, A. M., Dorian, K. T., Vaughan, H. A., Davis, I. D., Silvapulle, M., & Cebon, J. S. (2000). MAGE-12 and MAGE-6 are frequently expressed in malignant melanoma. Melanoma Research, 10(3), 259–264.

    Article  CAS  PubMed  Google Scholar 

  • Gong, M., Meng, L., Jiang, B., Zhang, J., Yang, H., Wu, J., & Shou, C. (2008). p37 from mycoplasma hyorhinis promotes cancer cell invasiveness and metastasis through activation of MMP-2 and followed by phosphorylation of EGFR. Molecular Cancer Therapeutics, 7(3), 530–537.

    Article  CAS  PubMed  Google Scholar 

  • Huang da, W., Sherman, B. T., & Lempicki, R. A. (2009). Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nature Protocols, 4(1), 44–57.

    Article  PubMed  Google Scholar 

  • Jiang, S., Zhang, S., Langenfeld, J., Lo, S. C., & Rogers, M. B. (2008). Mycoplasma infection transforms normal lung cells and induces bone morphogenetic protein 2 expression by post-transcriptional mechanisms. Journal of Cellular Biochemistry, 104(2), 580–594.

    Article  CAS  PubMed  Google Scholar 

  • Kirchhoff, H., Maass, C., Runge, M., Franz, B., Schmidt, R., Quentmeier, H., & Muhlradt, P. F. (1992). Tetrazolium [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] reduction by mycoplasmas. International Journal of Systematic Bacteriology, 42(3), 506–508.

    Article  CAS  PubMed  Google Scholar 

  • Kondo, S., Wakisaka, N., Muramatsu, M., Zen, Y., Endo, K., Murono, S., & Yoshizaki, T. (2011). Epstein-barr virus latent membrane protein 1 induces cancer stem/progenitor-like cells in nasopharyngeal epithelial cell lines. Journal of Virology, 85(21), 11255–11264. doi:10.1128/jvi.00188-11.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Langdon, W. B. (2014). Mycoplasma contamination in the 1000 genomes project. BioData Mining, 7, 3. doi:10.1186/1756-0381-7-3.

    Article  PubMed Central  PubMed  Google Scholar 

  • Liu, W. B., Zhang, J. Z., Jiang, B. H., Ren, T. T., Gong, M. M., Meng, L., & Shou, C. C. (2006). Lipoprotein p37 from mycoplasma hyorhinis inhibiting mammalian cell adhesion. Journal of Biomedical Science, 13(3), 323–331.

    Article  CAS  PubMed  Google Scholar 

  • Logunov, D. Y., Scheblyakov, D. V., Zubkova, O. V., Shmarov, M. M., Rakovskaya, I. V., Gurova, K. V, Gudkov, A. V. (2008). Mycoplasma infection suppresses p53, activates NF-kappaB and cooperates with oncogenic ras in rodent fibroblast transformation. Oncogene, 27(33), 4521–4531.

  • Mariotti, E., Gemei, M., Mirabelli, P., D’Alessio, F., Di Noto, R., Fortunato, G., & Del Vecchio, L. (2010). The percentage of CD133+ cells in human colorectal cancer cell lines is influenced by mycoplasma hyorhinis infection. BMC Cancer, 10(1), 120.

    Article  PubMed Central  PubMed  Google Scholar 

  • McLaughlin-Drubin, M. E., Crum, C. P., & Munger, K. (2011). Human papillomavirus E7 oncoprotein induces KDM6A and KDM6B histone demethylase expression and causes epigenetic reprogramming. Proceedings of the National Academy of Sciences of the United States of America, 108(5), 2130–2135. doi:10.1073/pnas.1009933108.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Miller, C. J., Kassem, H. S., Pepper, S. D., Hey, Y., Ward, T. H., & Margison, G. P. (2003). Mycoplasma infection significantly alters microarray gene expression profiles. Biotechniques, 35(4), 812–814.

    CAS  PubMed  Google Scholar 

  • Pollard, S. M., Yoshikawa, K., Clarke, I. D., Danovi, D., Stricker, S., Russell, R., & Dirks, P. (2009). Glioma stem cell lines expanded in adherent culture have tumor-specific phenotypes and are suitable for chemical and genetic screens. Cell Stem Cell, 4(6), 568–580. doi:10.1016/j.stem.2009.03.014.

    Article  CAS  PubMed  Google Scholar 

  • Romorini, L., Riva, D. A., Blüguermann, C., Videla Richardson, G. A., Scassa, M. E., Sevlever, G. E., & Miriuka, S. G. (2013). Effect of Antibiotics against Mycoplasma sp. on Human Embryonic Stem Cells Undifferentiated Status, Pluripotency, Cell Viability and Growth. PloS One, 8(7), e70267. doi:10.1371/journal.pone.0070267.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • van Staveren, W. C., Solis, D. Y., Hebrant, A., Detours, V., Dumont, J. E., & Maenhaut, C. (2009). Human cancer cell lines: experimental models for cancer cells in situ? For cancer stem cells? Biochimica et Biophysica Acta, 1795(2), 92–103.

    PubMed  Google Scholar 

  • Timenetsky, J., Santos, L. M., Buzinhani, M., & Mettifogo, E. (2006). Detection of multiple mycoplasma infection in cell cultures by PCR. Brazilian Journal of Medical and Biological Research, 39(7), 907–914.

    Article  CAS  PubMed  Google Scholar 

  • Uphoff, C. C., & Drexler, H. G. (2005a). Detection of mycoplasma contaminations. Methods in Molecular Biology, 290, 13–23.

    CAS  PubMed  Google Scholar 

  • Uphoff, C. C., & Drexler, H. G. (2005b). Eradication of mycoplasma contaminations. Methods in Molecular Biology, 290, 25–34.

    CAS  PubMed  Google Scholar 

  • Wong-Lee, J. G., Lovett, M. (1993). Rapid and Sensitive PCR Method for Identification of Mycoplasma Species in Tissue Culture. In: Persing D, Smith TF, Tenover FC (eds) Diagnostic Molecular and Microbiology: Principles and Applications. (pp. 257–260). Washington, DC: American Society for Microbiology.

Download references

Acknowledgments

CG was kindly supported by an NHMRC Biomedical Research PhD Scholarship (280925), a MOGA/COSA/Roche HOTT Fellowship and a Royal Australasian College of Physicians Australia Post Fellowship. MA was supported by a Cancer Council of Victoria PhD Scholarship. JC was a Practitioner Fellow of the National Health and Medical Research Council (NHMRC; 487905). IDD was supported by a Victorian Cancer Agency Clinician Researcher Fellowship and was an NHMRC Practitioner Fellow (487907). We acknowledge the support of a Melanoma Research Alliance (MRA) Team science Award and the Victorian State Government Operational Infrastructure Support Program for partial funding of this project.

Author Contributions

Craig Gedye: Conception and design, collection and/or assembly of data, analysis and interpretation, manuscript writing, final approval of manuscript.

Tracy Cardwell: Collection and/or assembly of data.

Nektaria Dimopoulos: Collection and/or assembly of data.

Bee Shin Tan: Collection and/or assembly of data.

Heather Jackson: Collection and/or assembly of data.

Suzanne Svobodová: Collection and/or assembly of data.

Matthew Anaka: Collection and/or assembly of data, data analysis and interpretation.

Andreas Behren: Collection and/or assembly of data, data analysis and interpretation.

Christopher Maher: data analysis and interpretation.

Oliver Hofmann: data analysis and interpretation.

Winston Hide: Conception and design, provision of study materials.

Otavia Caballero: Conception and design, provision of study materials, final approval of manuscript.

Ian D. Davis: Conception and design, provision of study materials, final approval of manuscript.

Jonathan Cebon: Conception and design, financial support, provision of study materials, final approval of manuscript.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Jonathan Cebon.

Ethics declarations

Conflict of Interest

We declare that we have no potential conflicts of interest in this work.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Gedye, C., Cardwell, T., Dimopoulos, N. et al. Mycoplasma Infection Alters Cancer Stem Cell Properties in Vitro. Stem Cell Rev and Rep 12, 156–161 (2016). https://doi.org/10.1007/s12015-015-9630-8

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12015-015-9630-8

Keywords

Navigation