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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...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Company of Biologists
2015
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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. |
format | Online Article Text |
id | pubmed-4582177 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | The Company of Biologists |
record_format | MEDLINE/PubMed |
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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