Expansion of CD133(+) colon cancer cultures retaining stem cell properties to enable cancer stem cell target discovery

Br J Cancer. 2010 Apr 13;102(8):1265-75. doi: 10.1038/sj.bjc.6605610. Epub 2010 Mar 23.


Background: Despite earlier studies demonstrating in vitro propagation of solid tumour cancer stem cells (CSCs) as non-adherent tumour spheres, it remains controversial as to whether CSCs can be maintained in vitro. Additional validation of the CSC properties of tumour spheres would support their use as CSC models and provide an opportunity to discover additional CSC cell surface markers to aid in CSC detection and potential elimination.

Methods: Primary tumour cells isolated from 13 surgically resected colon tumour specimens were propagated using serum-free CSC-selective conditions. The CSC properties of long-term cultured tumour spheres were established and mass spectrometry-based proteomics performed.

Results: Freshly isolated CD133(+) colorectal cancer cells gave rise to long-term tumour sphere (or spheroids) cultures maintaining CD133 expression. These spheroid cells were able to self-renew and differentiate into adherent epithelial lineages and recapitulate the phenotype of the original tumour. Relative to their differentiated progeny, tumour spheroid cells were more resistant to the chemotherapeutic irinotecan. Finally, CD44, CD166, CD29, CEACAM5, cadherin 17, and biglycan were identified by mass spectrometry to be enriched in CD133(+) tumour spheroid cells.

Conclusion: Our data suggest that ex vivo-expanded colon CSCs isolated from clinical specimens can be maintained in culture enabling the identification of CSC cell surface-associated proteins.

Publication types

  • Validation Study

MeSH terms

  • AC133 Antigen
  • ATP Binding Cassette Transporter, Subfamily G, Member 2
  • ATP-Binding Cassette Transporters / genetics
  • Adult
  • Aged
  • Aged, 80 and over
  • Animals
  • Antigens, CD / metabolism*
  • Cell Differentiation
  • Colonic Neoplasms / pathology*
  • Drug Resistance, Neoplasm
  • Female
  • Glycoproteins / metabolism*
  • Humans
  • Male
  • Mass Spectrometry
  • Membrane Proteins / analysis
  • Mice
  • Mice, Nude
  • Mice, SCID
  • Middle Aged
  • Neoplasm Proteins / genetics
  • Neoplasm Transplantation
  • Neoplastic Stem Cells*
  • Peptides / metabolism*
  • Proteomics
  • Spheroids, Cellular*
  • Tumor Cells, Cultured*


  • ABCG2 protein, human
  • AC133 Antigen
  • ATP Binding Cassette Transporter, Subfamily G, Member 2
  • ATP-Binding Cassette Transporters
  • Antigens, CD
  • Glycoproteins
  • Membrane Proteins
  • Neoplasm Proteins
  • PROM1 protein, human
  • Peptides
  • Prom1 protein, mouse