Imaging and analysis of 3D tumor spheroids enriched for a cancer stem cell phenotype

Fredika M. Robertson, Marcia A. Ogasawara, Ye Zaiming Ye, Khoi Chu, Ross Pickei, Bisrat G. Debeb, Wendy A. Woodward, Walter N. Hittelman, Massimo Cristofanilli, Sanford H. Barsky

Research output: Contribution to journalArticlepeer-review

43 Scopus citations


Tumors that display a highly metastatic phenotype contain subpopulations of cells that display characteristics similar to embryonic stem cells. These cells exhibit the ability to undergo self-renewal; slowly replicate to retain a nucleoside analog label, leading to their definition as "label-retaining cells"; express specific surface markers such as CD44+/CD24 -/low and CD133; and can give rise to cells of different lineages (i.e., they exhibit multipotency). Based on these characteristics, as well as their demonstrated ability to give rise to tumors in vivo, these cells have been defined as tumor-initiating cells (TICs), tumor-propagating cells, or cancer stem cells (CSCs). These cells are highly resistant to chemotherapeutic agents and radiation and are believed to be responsible for the development of both primary tumors and metastatic lesions at sites distant from the primary tumor. Established cancer cell lines contain CSCs, which can be propagated in vitro using defined conditions, to form 3D tumor spheroids. Because the vast majority of studies to identify cancer-associated genes and therapeutic targets use adherent cells grown in 2 dimensions on a plastic substrate, the multicellular composition of these 3D tumor spheroids presents both challenges and opportunities for their imaging and characterization. The authors describe approaches to image and analyze the properties of CSCs within 3D tumor spheroids, which can serve as the basis for defining the gene and protein signatures of CSCs and to develop therapeutic strategies that will effectively target this critically important population of cells that may be responsible for tumor progression.

Original languageEnglish (US)
Pages (from-to)820-829
Number of pages10
JournalJournal of Biomolecular Screening
Issue number7
StatePublished - Aug 2010


  • CyGEL™
  • EdU
  • cancer stem cells
  • click chemistry
  • confocal microscopy
  • imaging
  • inflammatory breast cancer
  • label-retaining cells
  • thermo-reversible cell mountant
  • tumor spheroids

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biotechnology
  • Biochemistry
  • Molecular Medicine
  • Pharmacology
  • Drug Discovery

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