Isolation of Cancer Stem Cells from Squamous Cell Carcinoma

  • Silvia FonteneteEmail author
  • Mirna Perez-MorenoEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1879)


Different cancer stem cell (CSC) populations can be found in many types of cancer, including squamous cell carcinoma (SSC). Diverse reports showed that CSC play a crucial role in the relapse of different types of cancer. CSC sustains tumor growth due to their capacity to self-renew and their potential to initiate secondary tumors with metastatic cancer features. Therefore, the development of methods for the isolation of CSC is a key step to explore the mechanisms underlying CSC maintenance. In this chapter, we provide a method for isolating CSC from cutaneous SSC using immunofluorescence labeling to allow the specific purification of CSC by fluorescence-activated cell sorting (FACS). This method is based on the use of CSC membrane markers, allowing as well the isolation CSC from different mouse strains.


Cancer stem cells FACS Mouse models Skin Squamous cell carcinoma 



The Perez-Moreno lab is supported by grants from the Worldwide Cancer Research UK Foundation (15-1219) and the Novo Nordisk Foundation (NNF17OC0028028).


  1. 1.
    Eisemann N, Waldmann A, Geller AC, Weinstock MA, Volkmer B, Greinert R, Breitbart EW, Katalinic A (2014) Non-melanoma skin cancer incidence and impact of skin cancer screening on incidence. J Invest Dermatol 134(1):43–50. Scholar
  2. 2.
    Lapouge G, Youssef KK, Vokaer B, Achouri Y, Michaux C, Sotiropoulou PA, Blanpain C (2011) Identifying the cellular origin of squamous skin tumors. Proc Natl Acad Sci U S A 108(18):7431–7436. Scholar
  3. 3.
    White AC, Tran K, Khuu J, Dang C, Cui Y, Binder SW, Lowry WE (2011) Defining the origins of Ras/p53-mediated squamous cell carcinoma. Proc Natl Acad Sci U S A 108(18):7425–7430. Scholar
  4. 4.
    Driessens G, Beck B, Caauwe A, Simons BD, Blanpain C (2012) Defining the mode of tumour growth by clonal analysis. Nature 488(7412):527–530. Scholar
  5. 5.
    Schepers AG, Snippert HJ, Stange DE, van den Born M, van Es JH, van de Wetering M, Clevers H (2012) Lineage tracing reveals Lgr5+ stem cell activity in mouse intestinal adenomas. Science 337(6095):730–735. Scholar
  6. 6.
    Morris RJ, Bortner CD, Cotsarelis G, Reece JM, Trempus CS, Faircloth RS, Tennant RW (2003) Enrichment for living murine keratinocytes from the hair follicle bulge with the cell surface marker CD34. J Invest Dermatol 120(4):501–511. Scholar
  7. 7.
    Trempus CS, Morris RJ, Ehinger M, Elmore A, Bortner CD, Ito M, Cotsarelis G, Nijhof JGW, Peckham J, Flagler N, Kissling G, Humble MM, King LC, Adams LD, Desai D, Amin S, Tennant RW (2007) CD34 expression by hair follicle stem cells is required for skin tumor development in mice. Cancer Res 67(9):4173–4181. Scholar
  8. 8.
    Schober M, Fuchs E (2011) Tumor-initiating stem cells of squamous cell carcinomas and their control by TGF-β and integrin/focal adhesion kinase (FAK) signaling. Proc Natl Acad Sci U S A 108(26):10544–10549. Scholar
  9. 9.
    Blanpain C, Lowry WE, Geoghegan A, Polak L, Fuchs E (2004) Self-renewal, multipotency, and the existence of two cell populations within an epithelial stem cell niche. Cell 118(5):635–648. Scholar
  10. 10.
    Lapouge G, Beck B, Nassar D, Dubois C, Dekoninck S, Blanpain C (2012) Skin squamous cell carcinoma propagating cells increase with tumour progression and invasiveness. EMBO J 31(24):4563–4575. Scholar
  11. 11.
    Terris B, Cavard C, Perret C (2010) EpCAM, a new marker for cancer stem cells in hepatocellular carcinoma. J Hepatol 52(2):280–281. Scholar
  12. 12.
    Osta WA, Chen Y, Mikhitarian K, Mitas M, Salem M, Hannun YA, Cole DJ, Gillanders WE (2004) EpCAM is overexpressed in breast cancer and is a potential target for breast cancer gene therapy. Cancer Res 64(16):5818–5824. Scholar
  13. 13.
    Gebauer F, Struck L, Tachezy M, Vashist Y, Wicklein D, Schumacher U, Izbicki JR, Bockhorn M (2014) Serum EpCAM expression in pancreatic cancer. Anticancer Res 34(9):4741–4746PubMedGoogle Scholar
  14. 14.
    Nowak JA, Fuchs E (2009) Isolation and culture of epithelial stem cells. Methods Mol Biol 482:215–232. Scholar

Copyright information

© Springer Science+Business Media New York 2018

Authors and Affiliations

  1. 1.Section for Cell Biology and Physiology, Department of Biology, Faculty of ScienceUniversity of CopenhagenCopenhagen ØDenmark

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