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. 2015 Apr 7;15:228. doi: 10.1186/s12885-015-1235-8

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© Barbieri et al.; licensee BioMed Central. 2015

This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Morphological appearance, proliferation and phenotyiping of CMC cultures. a) Morphological changes of floating mammospheres/clusters in stem cell-permissive medium (upper panels) from a cobblestone-like morphology to spindle-like cells in adherent monolayers (lower panels), after differentiation for 15 days in serum-containing medium. Phase-contrast images, original magnification 10X. b) Representative growth curves of CMC cells selected under stem cell-permissive or differentiation medium, showing the in vitro proliferative potential of cultures. Arbitrary units (a.u.) are referred to the number of living cells at day 1. Data represent the mean ± s.e.m. c) Enrichment in CD44⁺ cells and marker expression profile of CMC cells grown in stem cell-permissive or differentiation medium. Immunofluorescence analysis of pan-cytokeratin (Pan-CK), EGFR, ER-α, CXCR4 and CD44 in CMC spheroids (upper panels) and after 15-day exposure to serum-containing medium. Images from confocal microscopy, original magnification 100X.