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Quantification of regenerative potential in primary human mammary epithelial cells

We present an organoid regeneration assay in which freshly isolated human mammary epithelial cells are cultured in adherent or floating collagen gels, corresponding to a rigid or compliant matrix environment. In both conditions, luminal progenitors form spheres, whereas basal cells generate branched...

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Autores principales: Linnemann, Jelena R., Miura, Haruko, Meixner, Lisa K., Irmler, Martin, Kloos, Uwe J., Hirschi, Benjamin, Bartsch, Harald S., Sass, Steffen, Beckers, Johannes, Theis, Fabian J., Gabka, Christian, Sotlar, Karl, Scheel, Christina H.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Company of Biologists 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4582177/
https://www.ncbi.nlm.nih.gov/pubmed/26071498
http://dx.doi.org/10.1242/dev.123554
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author Linnemann, Jelena R.
Miura, Haruko
Meixner, Lisa K.
Irmler, Martin
Kloos, Uwe J.
Hirschi, Benjamin
Bartsch, Harald S.
Sass, Steffen
Beckers, Johannes
Theis, Fabian J.
Gabka, Christian
Sotlar, Karl
Scheel, Christina H.
author_facet Linnemann, Jelena R.
Miura, Haruko
Meixner, Lisa K.
Irmler, Martin
Kloos, Uwe J.
Hirschi, Benjamin
Bartsch, Harald S.
Sass, Steffen
Beckers, Johannes
Theis, Fabian J.
Gabka, Christian
Sotlar, Karl
Scheel, Christina H.
author_sort Linnemann, Jelena R.
collection PubMed
description We present an organoid regeneration assay in which freshly isolated human mammary epithelial cells are cultured in adherent or floating collagen gels, corresponding to a rigid or compliant matrix environment. In both conditions, luminal progenitors form spheres, whereas basal cells generate branched ductal structures. In compliant but not rigid collagen gels, branching ducts form alveoli at their tips, express basal and luminal markers at correct positions, and display contractility, which is required for alveologenesis. Thereby, branched structures generated in compliant collagen gels resemble terminal ductal-lobular units (TDLUs), the functional units of the mammary gland. Using the membrane metallo-endopeptidase CD10 as a surface marker enriches for TDLU formation and reveals the presence of stromal cells within the CD49f(hi)/EpCAM(−) population. In summary, we describe a defined in vitro assay system to quantify cells with regenerative potential and systematically investigate their interaction with the physical environment at distinct steps of morphogenesis.
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spelling pubmed-45821772015-11-04 Quantification of regenerative potential in primary human mammary epithelial cells Linnemann, Jelena R. Miura, Haruko Meixner, Lisa K. Irmler, Martin Kloos, Uwe J. Hirschi, Benjamin Bartsch, Harald S. Sass, Steffen Beckers, Johannes Theis, Fabian J. Gabka, Christian Sotlar, Karl Scheel, Christina H. Development Techniques and Resources We present an organoid regeneration assay in which freshly isolated human mammary epithelial cells are cultured in adherent or floating collagen gels, corresponding to a rigid or compliant matrix environment. In both conditions, luminal progenitors form spheres, whereas basal cells generate branched ductal structures. In compliant but not rigid collagen gels, branching ducts form alveoli at their tips, express basal and luminal markers at correct positions, and display contractility, which is required for alveologenesis. Thereby, branched structures generated in compliant collagen gels resemble terminal ductal-lobular units (TDLUs), the functional units of the mammary gland. Using the membrane metallo-endopeptidase CD10 as a surface marker enriches for TDLU formation and reveals the presence of stromal cells within the CD49f(hi)/EpCAM(−) population. In summary, we describe a defined in vitro assay system to quantify cells with regenerative potential and systematically investigate their interaction with the physical environment at distinct steps of morphogenesis. The Company of Biologists 2015-09-15 /pmc/articles/PMC4582177/ /pubmed/26071498 http://dx.doi.org/10.1242/dev.123554 Text en © 2015. Published by The Company of Biologists Ltd http://creativecommons.org/licenses/by/3.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.
spellingShingle Techniques and Resources
Linnemann, Jelena R.
Miura, Haruko
Meixner, Lisa K.
Irmler, Martin
Kloos, Uwe J.
Hirschi, Benjamin
Bartsch, Harald S.
Sass, Steffen
Beckers, Johannes
Theis, Fabian J.
Gabka, Christian
Sotlar, Karl
Scheel, Christina H.
Quantification of regenerative potential in primary human mammary epithelial cells
title Quantification of regenerative potential in primary human mammary epithelial cells
title_full Quantification of regenerative potential in primary human mammary epithelial cells
title_fullStr Quantification of regenerative potential in primary human mammary epithelial cells
title_full_unstemmed Quantification of regenerative potential in primary human mammary epithelial cells
title_short Quantification of regenerative potential in primary human mammary epithelial cells
title_sort quantification of regenerative potential in primary human mammary epithelial cells
topic Techniques and Resources
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4582177/
https://www.ncbi.nlm.nih.gov/pubmed/26071498
http://dx.doi.org/10.1242/dev.123554
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