TY - JOUR
T1 - Culture and characterization of mammary cancer stem cells in mammospheres
AU - Piscitelli, Eleonora
AU - Cocola, Cinzia
AU - Thaden, Frank Rüdiger
AU - Pelucchi, Paride
AU - Gray, Brian
AU - Bertalot, Giovanni
AU - Albertini, Alberto
AU - Reinbold, Rolland
AU - Zucchi, Ileana
PY - 2014
Y1 - 2014
N2 - 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.
AB - 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.
KW - ALDEFLUOR
KW - Cancer stem cells
KW - CellVue Maroon dye
KW - LA7CSCs
KW - Mammospheres
KW - Stem cells
KW - Tubules and cysts
UR - http://www.scopus.com/inward/record.url?scp=84921733972&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84921733972&partnerID=8YFLogxK
U2 - 10.1007/978-1-4939-1785-3_18
DO - 10.1007/978-1-4939-1785-3_18
M3 - Article
C2 - 25388398
AN - SCOPUS:84921733972
VL - 1235
SP - 243
EP - 262
JO - Methods in Molecular Biology
JF - Methods in Molecular Biology
SN - 1064-3745
ER -