Culture and characterization of mammary cancer stem cells in mammospheres

Eleonora Piscitelli, Cinzia Cocola, Frank Rüdiger Thaden, Paride Pelucchi, Brian Gray, Giovanni Bertalot, Alberto Albertini, Rolland Reinbold, Ileana Zucchi

Research output: Contribution to journalArticlepeer-review


Mammospheres (MMs) are a model for culturing and maintaining mammary gland stem cells (SCs) or cancer stem cells (CSCs) ex situ. As MMs recapitulate the micro-niche of the mammary gland or a tumor, MMs are a model for studying the properties of SCs or CSCs, and for mapping, isolating, and characterizing the SC/CSC generated lineages. Cancer stem cells share with normal SCs the properties of selfrenewal and the capacity to generate all cell types and organ structures of the mammary gland. Analysis of human tumor samples suggests that CSCs are heterogeneous in terms of proliferation and differentiation potential. Mammospheres from CSCs likewise display heterogeneity. This heterogeneity makes analysis of CSC generated MMs challenging. To identify the unique and diverse properties of MM derived CSCs, comparative analysis with MMs obtained from normal SCs is required. Here we present protocols for identifying and enriching cells with SC features from a cancer cell line using the LA7CSCs as a model. A comprehensive and comparative approach for identifying, isolating, and characterizing MMs from SCs and CSCs from human breast is also introduced. In addition, we describe detailed procedures for identifying, isolating, and characterizing mammary gland specific cell types, generated during MM formation.

Original languageEnglish
Pages (from-to)243-262
Number of pages20
JournalMethods in molecular biology (Clifton, N.J.)
Publication statusPublished - 2014


  • Cancer stem cells
  • CellVue Maroon dye
  • LA7CSCs
  • Mammospheres
  • Stem cells
  • Tubules and cysts

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Medicine(all)


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