Abstract
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.
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Acknowledgements and Financial Support
We like to acknowledge the expert technical assistance of Sig. Emanuele Canonico of the FRACTAL-Servizio di Citometria Istituto Scientifico San Raffaele, Milano and the always supportive Loredana Ansalone for her administrative help. We thank the support Dr. Martin Götte from the Universitätsklinikum of Münster, Germany and the staff of the Department of Pathology of Leno (BS) for their excellent technical support. We acknowledge and are grateful to the IZ grant support: the N.O.B.E.L. Grant and Cariplo Progetti-Internazionali n. 2008–2015 (Fondazione CARIPLO), MIUR-FIRB Newton grant RBAP11BYNP and Fierce grant RBAP11BYNP. R.A.R is supported with a fellowship from the Fondazione CARIPLO Progetti-Internazionali. This work is dedicated to the memory of a dear friend, Sabina Cascella who died of cancer when she was still young and full of life.
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Piscitelli, E. et al. (2015). Culture and Characterization of Mammary Cancer Stem Cells in Mammospheres. In: Rich, I. (eds) Stem Cell Protocols. Methods in Molecular Biology, vol 1235. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1785-3_18
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DOI: https://doi.org/10.1007/978-1-4939-1785-3_18
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-1785-3
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