In vitro tumorigenic assay: colony forming assay for cancer stem cells

Vijayalakshmi Rajendran, Mayur Vilas Jain

Research output: Chapter in Book/Report/Conference proceedingBook chapterResearchpeer-review


Colony forming or clonogenic assay is an in vitro quantitative technique to examine the capability of a single cell to grow into a large colony through clonal expansion. Clonogenic activity is a sensitive indicator of undifferentiated cancer stem cells. Here, we described the colony forming ability of the isolated breast cancer stem cells from the total population of cancer cells using double-layered, soft agarose-based assay. This method demonstrates that cancer stem cells can survive and generate colony growth in an anchorage-independent culture model. The 0.005% crystal violet solution is used in this assay to visualize the generated colonies.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press
Number of pages7
Publication statusPublished - 2018

Publication series

NameMethods in Molecular Biology
ISSN (Print)10643745

Subject classification (UKÄ)

  • Cell and Molecular Biology
  • Cancer and Oncology

Free keywords

  • Agarose
  • Cancer stem cell
  • Clonogenic assay
  • Colony growth
  • Crystal violet


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