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Long-term cultures of stem/progenitor cells from lobular and ductal breast carcinomas under non-adherent conditions

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Abstract

A small subpopulation of stem/progenitor cells can give rise to the diversity of differentiated cells that comprise the bulk of the tumor. Are proliferating cells, within the bulk of tumor, few cells with uncommon features? The cell biological approach provides a limitless model for studying the hierarchical organization of progenitor subpopulation and identifying potential therapeutic targets. Aim of the study was to expand patients’ breast cancer cells for evaluating functional cell properties, and to characterize the protein expression profile of selected cells to be compared with that of primary tumors. Breast cancer cells from estrogen receptor (ERα) positive, HER2 negative lobular (LoBS cells) and ductal (DuBS cells) histotype were cultured under non-adherent conditions to form mammospheres. Sorting of the cells by their surface expression of CD24 and CD44 gave rise to subpopulations which were propagated, enriched and characterized for the expression of epithelial and stromal markers. We found that non-adherent culture conditions generate mammospheres of slowly proliferating cells; single cells, dissociated from mammospheres, grow in soft agar; long-term cultured LoBS and DuBS cells, CD44+/CD24low, express cytokeratin 5 (CK5), α-smooth muscle actin (α-sma) and vimentin, known as markers of basal/myoepithelial cells; and ERα (only DuBS cells), HER1 (EGF-Receptor), activated HER2, and cyclinD1 as markers of luminal epithelial cell. Isolates of cells from breast cancer patients may be a tool for a marker-driven testing of targeted therapies.

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Acknowledgments

We declare no conflict of interest. This work was supported by: Associazione Italiana per la Ricerca sul Cancro, CRPO Centro Regionale di Prevenzione Oncologica, Regione Campania, Ministero dell’Universita` e Ricerca, and Ministero della Salute, Italia.

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Authors and Affiliations

  1. Dipartimento di Biologia e Patologia Cellulare e Molecolare “L. Califano”, Università di Napoli Federico II, via Pansini 5, 80131, Naples, Italy

    Agostina Nardone, Sara Corvigno, Annalisa Brescia, Daniel D’Andrea & Bianca Maria Veneziani

  2. Dipartimento di Chirurgia Generale Oncologica e Tecnologie Avanzate, Università degli Studi di Napoli Federico II, via Pansini 5, 80131, Naples, Italy

    Gennaro Limite

  3. Oncotech, Facoltà di Medicina e Chirurgia, Università degli Studi di Napoli Federico II, via Pansini 5, 80131, Naples, Italy

    Bianca Maria Veneziani

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  1. Agostina Nardone
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  2. Sara Corvigno
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  3. Annalisa Brescia
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  4. Daniel D’Andrea
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  5. Gennaro Limite
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  6. Bianca Maria Veneziani
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Correspondence to Bianca Maria Veneziani.

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A. Nardone and S. Corvigno contributed equally to this work.

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Nardone, A., Corvigno, S., Brescia, A. et al. Long-term cultures of stem/progenitor cells from lobular and ductal breast carcinomas under non-adherent conditions. Cytotechnology 63, 67–80 (2011). https://doi.org/10.1007/s10616-010-9328-3

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