Isolation and Propagation of Colon Cancer Stem Cells

  • Pramudita R. Prasetyanti
  • Cheryl Zimberlin
  • Felipe De Sousa E. Melo
  • Jan Paul Medema
Part of the Methods in Molecular Biology book series (MIMB, volume 1035)


The design of tissue culture conditions that faithfully reproduce the characteristics of cells in their native environment remains one of the main challenges of cancer stem cell (CSC) biology. Here we describe a detailed methodology for the isolation and expansion of both human colon CSCs and mouse intestinal adenoma together with a brief differentiation and coculture method that proved to be valuable to study the concept of CSCs plasticity.

Key words

Colorectal cancer Adenoma Cancer stem cells CD133 Wnt signaling Ultralow adherent EGF Isolation 


  1. 1.
    Clarke MF, Dick JE, Dirks PB, Eaves CJ, Jamieson CH et al (2006) Cancer stem cells–perspectives on current status and future directions: AACR Workshop on cancer stem cells. Cancer Res 66:9339–9344PubMedCrossRefGoogle Scholar
  2. 2.
    Huelsken MM, Wakamiya J, Ding J, van de Wetering M, Clevers H et al (2004) Beta-catenin regulates Cripto expression during embryogenesis and tumor formation. Eur J Cell Biol 83:31Google Scholar
  3. 3.
    Lee J, Kotliarova S, Kotliarov Y, Li A, Su Q et al (2006) Tumor stem cells derived from glioblastomas cultured in bFGF and EGF more closely mirror the phenotype and genotype of primary tumors than do serum-cultured cell lines. Cancer Cell 9:391–403PubMedCrossRefGoogle Scholar
  4. 4.
    O’Brien CA, Pollett A, Gallinger S, Dick JE (2007) A human colon cancer cell capable of initiating tumour growth in immunodeficient mice. Nature 445:106–110PubMedCrossRefGoogle Scholar
  5. 5.
    Ricci-Vitiani L, Lombardi DG, Pilozzi E, Biffoni M, Todaro M et al (2007) Identification and expansion of human colon-cancer-initiating cells. Nature 445:111–115PubMedCrossRefGoogle Scholar
  6. 6.
    Todaro M, Alea MP, Di Stefano AB, Cammareri P, Vermeulen L et al (2007) Colon cancer stem cells dictate tumor growth and resist cell death by production of interleukin-4. Cell Stem Cell 1:389–402PubMedCrossRefGoogle Scholar
  7. 7.
    Vermeulen L, De Sousa EM, van der Heijden M, Cameron K, de Jong JH et al (2010) Wnt activity defines colon cancer stem cells and is regulated by the microenvironment. Nat Cell Biol 12:468–476PubMedCrossRefGoogle Scholar
  8. 8.
    Vermeulen L, Todaro M, de Sousa MF, Sprick MR, Kemper K et al (2008) Single-cell cloning of colon cancer stem cells reveals a multi-lineage differentiation capacity. Proc Natl Acad Sci USA 105:13427–13432PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2013

Authors and Affiliations

  • Pramudita R. Prasetyanti
    • 1
  • Cheryl Zimberlin
    • 1
  • Felipe De Sousa E. Melo
    • 1
  • Jan Paul Medema
    • 1
  1. 1.Laboratory of Experimental Oncology and Radiobiology Center for Experimental Molecular MedicineAcademic Medical CenterAmsterdamThe Netherlands

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