Mapping mammary gland architecture using multi-scale in situ analysis (original) (raw)
Journal Article
Rodrigo Fernandez-Gonzalez
Department of Cancer Biology, Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA
Joint Graduate Group in Bioengineering, University of California, San Francisco/Berkeley, Berkeley, CA 94720, USA
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Department of Cancer Biology, Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA
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Department of Anatomy and the Biomedical Sciences Program, University of California, San Francisco, San Francisco, CA 94143, USA
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Department of Cancer Biology, Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA
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Department of Anatomy and the Biomedical Sciences Program, University of California, San Francisco, San Francisco, CA 94143, USA
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Carlos Ortiz-de-Solorzano
Morphology and Imaging Group and Cancer Imaging Laboratory, Center for Applied Medical Research, University of Navarre, Pamplona, 31008 Navarre, Spain
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Department of Cancer Biology, Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA
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Received:
29 September 2008
Accepted:
06 November 2008
Published:
05 December 2008
Cite
Rodrigo Fernandez-Gonzalez, Irineu Illa-Bochaca, Bryan E. Welm, Markus C. Fleisch, Zena Werb, Carlos Ortiz-de-Solorzano, Mary Helen Barcellos-Hoff, Mapping mammary gland architecture using multi-scale in situ analysis, Integrative Biology, Volume 1, Issue 1, January 2009, Pages 80–89, https://doi.org/10.1039/b816933k
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Abstract
We have built a novel computational microscopy platform that integrates image acquisition, storage, processing and analysis to study cell populations in situ. This platform allows high-content studies where multiple features are measured and linked at multiple scales. We used this approach to study the cellular composition and architecture of the mouse mammary gland by quantitatively tracking the distribution and type, position, proliferative state, and hormone receptor status of epithelial cells that incorporated bromodeoxyuridine while undergoing DNA synthesis during puberty and retained this label in the adult gland as a function of tissue structure. Immunofluorescence was used to identify label-retaining cells, as well as epithelial cells expressing the proteins progesterone receptor and P63. Only 3.6% of luminal cells were label-retaining cells, the majority of which did not express the progesterone receptor. Multi-scale in situ analysis revealed that luminal label-retaining cells have a distinct nuclear morphology, are enriched 3.4-fold in large ducts, and are distributed asymmetrically across the tissue. We postulated that LRC enriched in the ventral mammary gland represent progenitor cells. Epithelial cells isolated from the ventral versus the dorsal portion of the gland were enriched for the putative stem cell markers CD24 and CD49f as measured by fluorescence activated cell sorting. Thus, quantitative analysis of the cellular composition of the mammary epithelium across spatial scales identified a previously unrecognized architecture in which the ventral-most, large ducts contain a reservoir of undifferentiated, putative stem cells.
This journal is © The Royal Society of Chemistry 2009
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