Comparison of methods for the isolation of human breast epithelial and myoepithelial cells.

Two lineages, epithelial, and myoepithelial cells are the main cell populations in the normal mammary gland and in breast cancer. Traditionally, cancer research has been performed using commercial cell lines, but primary cell cultures obtained from fresh breast tissue are a powerful tool to study mo...

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Autores: Zubeldia Plazaola, Arantzazu, Ametller, Elisabet, Mancino, Mario, Prats de Puig, Miquel, López Plana, Anna, Guzman, Flavia, Vinyals, Laia, Pastor Arroyo, Eva M., Almendro Navarro, Vanessa, Fuster Orellana, Gemma, Gascón, Pere
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2015
País:España
Institución:Universidad de Barcelona
Repositorio:Dipòsit Digital de la UB
OAI Identifier:oai:diposit.ub.edu:2445/120275
Acceso en línea:https://hdl.handle.net/2445/120275
Access Level:acceso abierto
Palabra clave:Cèl·lules epitelials
Càncer de mama
Glàndules mamàries
Oncologia
Epithelial cells
Breast cancer
Mammary glands
Oncology
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spelling Comparison of methods for the isolation of human breast epithelial and myoepithelial cells.Zubeldia Plazaola, ArantzazuAmetller, ElisabetMancino, MarioPrats de Puig, MiquelLópez Plana, AnnaGuzman, FlaviaVinyals, LaiaPastor Arroyo, Eva M.Almendro Navarro, VanessaFuster Orellana, GemmaGascón, PereCèl·lules epitelialsCàncer de mamaGlàndules mamàriesOncologiaEpithelial cellsBreast cancerMammary glandsOncologyTwo lineages, epithelial, and myoepithelial cells are the main cell populations in the normal mammary gland and in breast cancer. Traditionally, cancer research has been performed using commercial cell lines, but primary cell cultures obtained from fresh breast tissue are a powerful tool to study more reliably new aspects of mammary gland biology, including normal and pathological conditions. Nevertheless, the methods described to date have some technical problems in terms of cell viability and yield, which hamper work with primary mammary cells. Therefore, there is a need to optimize technology for the proper isolation of epithelial and myoepithelial cells. For this reason, we compared four methods in an effort to improve the isolation and primary cell culture of different cell populations of human mammary epithelium. The samples were obtained from healthy tissue of patients who had undergone mammoplasty or mastectomy surgery. We based our approaches on previously described methods, and incorporated additional steps to ameliorate technical efficiency and increase cell survival. We determined cell growth and viability by phase-contrast images, growth curve analysis and cell yield, and identified cell-lineage specific markers by flow cytometry and immunofluorescence in 3D cell cultures. These techniques allowed us to better evaluate the functional capabilities of these two main mammary lineages, using CD227/K19 (epithelial cells) and CD10/K14 (myoepithelial cells) antigens. Our results show that slow digestion at low enzymatic concentration combined with the differential centrifugation technique is the method that best fits the main goal of the present study: protocol efficiency and cell survival yield. In summary, we propose some guidelines to establish primary mammary epithelial cell lines more efficiently and to provide us with a strong research instrument to better understand the role of different epithelial cell types in the origin of breast cancer.Frontiers Media2015info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://hdl.handle.net/2445/120275Articles publicats en revistes (Medicina)reponame:Dipòsit Digital de la UBinstname:Universidad de BarcelonaInglésReproducció del document publicat a: https://doi.org/10.3389/fcell.2015.00032Frontiers in Cell and Developmental Biology, 2015, vol. 3, p. 32https://doi.org/10.3389/fcell.2015.00032cc-by (c) Zubeldia Plazaola, Arantzazu et al., 2015http://creativecommons.org/licenses/by/3.0/esinfo:eu-repo/semantics/openAccessoai:diposit.ub.edu:2445/1202752026-05-27T06:46:51Z
dc.title.none.fl_str_mv Comparison of methods for the isolation of human breast epithelial and myoepithelial cells.
title Comparison of methods for the isolation of human breast epithelial and myoepithelial cells.
spellingShingle Comparison of methods for the isolation of human breast epithelial and myoepithelial cells.
Zubeldia Plazaola, Arantzazu
Cèl·lules epitelials
Càncer de mama
Glàndules mamàries
Oncologia
Epithelial cells
Breast cancer
Mammary glands
Oncology
title_short Comparison of methods for the isolation of human breast epithelial and myoepithelial cells.
title_full Comparison of methods for the isolation of human breast epithelial and myoepithelial cells.
title_fullStr Comparison of methods for the isolation of human breast epithelial and myoepithelial cells.
title_full_unstemmed Comparison of methods for the isolation of human breast epithelial and myoepithelial cells.
title_sort Comparison of methods for the isolation of human breast epithelial and myoepithelial cells.
dc.creator.none.fl_str_mv Zubeldia Plazaola, Arantzazu
Ametller, Elisabet
Mancino, Mario
Prats de Puig, Miquel
López Plana, Anna
Guzman, Flavia
Vinyals, Laia
Pastor Arroyo, Eva M.
Almendro Navarro, Vanessa
Fuster Orellana, Gemma
Gascón, Pere
author Zubeldia Plazaola, Arantzazu
author_facet Zubeldia Plazaola, Arantzazu
Ametller, Elisabet
Mancino, Mario
Prats de Puig, Miquel
López Plana, Anna
Guzman, Flavia
Vinyals, Laia
Pastor Arroyo, Eva M.
Almendro Navarro, Vanessa
Fuster Orellana, Gemma
Gascón, Pere
author_role author
author2 Ametller, Elisabet
Mancino, Mario
Prats de Puig, Miquel
López Plana, Anna
Guzman, Flavia
Vinyals, Laia
Pastor Arroyo, Eva M.
Almendro Navarro, Vanessa
Fuster Orellana, Gemma
Gascón, Pere
author2_role author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Cèl·lules epitelials
Càncer de mama
Glàndules mamàries
Oncologia
Epithelial cells
Breast cancer
Mammary glands
Oncology
topic Cèl·lules epitelials
Càncer de mama
Glàndules mamàries
Oncologia
Epithelial cells
Breast cancer
Mammary glands
Oncology
description Two lineages, epithelial, and myoepithelial cells are the main cell populations in the normal mammary gland and in breast cancer. Traditionally, cancer research has been performed using commercial cell lines, but primary cell cultures obtained from fresh breast tissue are a powerful tool to study more reliably new aspects of mammary gland biology, including normal and pathological conditions. Nevertheless, the methods described to date have some technical problems in terms of cell viability and yield, which hamper work with primary mammary cells. Therefore, there is a need to optimize technology for the proper isolation of epithelial and myoepithelial cells. For this reason, we compared four methods in an effort to improve the isolation and primary cell culture of different cell populations of human mammary epithelium. The samples were obtained from healthy tissue of patients who had undergone mammoplasty or mastectomy surgery. We based our approaches on previously described methods, and incorporated additional steps to ameliorate technical efficiency and increase cell survival. We determined cell growth and viability by phase-contrast images, growth curve analysis and cell yield, and identified cell-lineage specific markers by flow cytometry and immunofluorescence in 3D cell cultures. These techniques allowed us to better evaluate the functional capabilities of these two main mammary lineages, using CD227/K19 (epithelial cells) and CD10/K14 (myoepithelial cells) antigens. Our results show that slow digestion at low enzymatic concentration combined with the differential centrifugation technique is the method that best fits the main goal of the present study: protocol efficiency and cell survival yield. In summary, we propose some guidelines to establish primary mammary epithelial cell lines more efficiently and to provide us with a strong research instrument to better understand the role of different epithelial cell types in the origin of breast cancer.
publishDate 2015
dc.date.none.fl_str_mv 2015
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv https://hdl.handle.net/2445/120275
url https://hdl.handle.net/2445/120275
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Reproducció del document publicat a: https://doi.org/10.3389/fcell.2015.00032
Frontiers in Cell and Developmental Biology, 2015, vol. 3, p. 32
https://doi.org/10.3389/fcell.2015.00032
dc.rights.none.fl_str_mv cc-by (c) Zubeldia Plazaola, Arantzazu et al., 2015
http://creativecommons.org/licenses/by/3.0/es
info:eu-repo/semantics/openAccess
rights_invalid_str_mv cc-by (c) Zubeldia Plazaola, Arantzazu et al., 2015
http://creativecommons.org/licenses/by/3.0/es
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Frontiers Media
publisher.none.fl_str_mv Frontiers Media
dc.source.none.fl_str_mv Articles publicats en revistes (Medicina)
reponame:Dipòsit Digital de la UB
instname:Universidad de Barcelona
instname_str Universidad de Barcelona
reponame_str Dipòsit Digital de la UB
collection Dipòsit Digital de la UB
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,300724

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