Culture and characterization of mammary cancer stem cells in mammospheres.
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 self-renewal 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.
Consiglio Nazionale delle Ricerche, Instituto di Technologie Biomediche, via Cervi 93, Segrate Milan, 20090, Italy.
Eleonora
E
Piscitelli
Cinzia
C
Cocola
Frank Rüdiger
FR
Thaden
Paride
P
Pelucchi
Brian
B
Gray
Giovanni
G
Bertalot
Alberto
A
Albertini
Rolland
R
Reinbold
Ileana
I
Zucchi
D000818:Animals; D001943:Breast Neoplasms / Q000473:pathology*; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D002469:Cell Separation / Q000379:methods; D005260:Female; D006801:Humans; D042361:Mammary Glands, Human / Q000166:cytology* / Q000473:pathology*; D014411:Neoplastic Stem Cells / Q000166:cytology* / Q000473:pathology*; D013194:Staining and Labeling / Q000379:methods