Isolation and Characterization of Cancer Stem Cells from Primary Head and Neck Squamous Cell Carcinoma Tumors

  • Hong S. Kim
  • Alexander T. Pearson
  • Jacques E. Nör
Protocol
First Online:
Part of the Methods in Molecular Biology book series (MIMB, volume 1395)

Abstract

Drug resistance remains a significant problem in the treatment of patients with head and neck squamous cell carcinoma (HNSCC). Recent reports showed that a subpopulation of highly tumorigenic cells, called cancer stem cells (CSCs), is uniquely resistant to chemotherapy, suggesting that these cells play an important role in the relapse of HNSCC. The development of methods for the isolation and culture of cancer stem cells is a key step to enable studies exploring the mechanisms underlying the role of these cells in chemoresistance. Here, we describe a method to isolate cancer stem cells from primary head and neck tumors and for the generation of orospheres.

Key words

Head and neck squamous cell carcinoma Cancer stem cells Chemoresistance Self-renewal Multipotency Orospheres 
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References

  1. 1.
    Kamangar F, Dores GM, Anderson WF (2006) Patterns of cancer incidence, mortality, and prevalence across five continents: defining priorities to reduce cancer disparities in different geographic regions of the world. J Clin Oncol 24:2137–2150CrossRefPubMedGoogle Scholar
  2. 2.
    Forastiere AA, Goepfert H, Maor M, Pajak TF, Weber R, Morrison W, Glisson B, Trotti A, Ridge JA, Chao C, Peters G, Lee DJ, Leaf A, Ensley J, Cooper J (2003) Concurrent chemotherapy and radiotherapy for organ preservation in advanced laryngeal cancer. N Engl J Med 349:2091–2098CrossRefPubMedGoogle Scholar
  3. 3.
    Pfister DG, Spencer S, Brizel DM, Burtness B, Busse PM, Caudell JJ, Cmelak AJ, Colevas AD, Dunphy F, Eisele DW, Gilbert J, Gillison ML, Haddad RI, Haughey BH, Hicks WL Jr, Hitchcock YJ, Jimeno A, Kies MS, Lydiatt WM, Maghami E, Martins R, McCaffrey T, Mell LK, Mittal BB, Pinto HA, Ridge JA, Rodriguez CP, Samant S, Schuller DE, Shah JP, Weber RS, Wolf GT, Worden F, Yom SS, McMillian NR, Hughes M (2014) Head and neck cancers, Version 2.2014. Clinical practice guidelines in oncology. J Natl Compr Canc Netw 12:1454–1487PubMedGoogle Scholar
  4. 4.
    Okamoto A, Chikamatsu K, Sakakura K, Hatsushika K, Takahashi G, Masuyama K (2009) Expansion and characterization of cancer stem-like cells in squamous cell carcinoma of the head and neck. Oral Oncol 45:633–639CrossRefPubMedGoogle Scholar
  5. 5.
    Zhang Q, Shi S, Yen Y, Brown J, Ta JQ, Le AD (2010) A subpopulation of CD133(+) cancer stem-like cells characterized in human oral squamous cell carcinoma confer resistance to chemotherapy. Cancer Lett 289:151–160CrossRefPubMedGoogle Scholar
  6. 6.
    Nor C, Zhang Z, Warner KA, Bernardi L, Visioli F, Helman JI, Roesler R, Nor JE (2014) Cisplatin induces Bmi-1 and enhances the stem cell fraction in head and neck cancer. Neoplasia 16:137–146CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Reers S, Pfannerstill AC, Maushagen R, Pries R, Wollenberg B (2014) Stem cell profiling in head and neck cancer reveals an Oct-4 expressing subpopulation with properties of chemoresistance. Oral Oncol 50:155–162CrossRefPubMedGoogle Scholar
  8. 8.
    Reynolds BA, Weiss S (1992) Generation of neurons and astrocytes from isolated cells of the adult mammalian central nervous system. Science 255:1707–1710CrossRefPubMedGoogle Scholar
  9. 9.
    Dontu G, Abdallah WM, Foley JM, Jackson KW, Clarke MF, Kawamura MJ, Wicha MS (2003) In vitro propagation and transcriptional profiling of human mammary stem/progenitor cells. Genes Dev 17:1253–1270CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Krishnamurthy S, Dong Z, Vodopyanov D, Imai A, Helman JI, Prince ME, Wicha MS, Nor JE (2010) Endothelial cell-initiated signaling promotes the survival and self-renewal of cancer stem cells. Cancer Res 70:9969–9978CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Krishnamurthy S, Nor JE (2013) Orosphere assay: a method for propagation of head and neck cancer stem cells. Head Neck 35:1015–1021CrossRefPubMedGoogle Scholar
  12. 12.
    Clay MR, Tabor M, Owen JH, Carey TE, Bradford CR, Wolf GT, Wicha MS, Prince ME (2010) Single-marker identification of head and neck squamous cell carcinoma cancer stem cells with aldehyde dehydrogenase. Head Neck 32:1195–1201CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Prince ME, Sivanandan R, Kaczorowski A, Wolf GT, Kaplan MJ, Dalerba P, Weissman IL, Clarke MF, Ailles LE (2007) Identification of a subpopulation of cells with cancer stem cell properties in head and neck squamous cell carcinoma. Proc Natl Acad Sci U S A 104:973–978CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Hong S. Kim
    • 1
  • Alexander T. Pearson
    • 2
    • 3
  • Jacques E. Nör
    • 1
    • 3
    • 4
    • 5
  1. 1.Department of Restorative SciencesUniversity of Michigan School of DentistryAnn ArborUSA
  2. 2.Division of Hematology/Oncology, Department of Internal MedicineUniversity of Michigan Medical CenterAnn ArborUSA
  3. 3.Comprehensive Cancer CenterUniversity of MichiganAnn ArborUSA
  4. 4.Department of Biomedical EngineeringUniversity of Michigan College of EngineeringAnn ArborUSA
  5. 5.Department of OtolaryngologyUniversity of Michigan School of MedicineAnn ArborUSA

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