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Microstructured soft devices for the growth and analysis of populations of homogenous multicellular tumor spheroids
Multicellular tumor spheroids are rapidly emerging as an improved in vitro model with respect to more traditional 2D culturing. Microwell culturing is a simple and accessible method for generating a large number of uniformly sized spheroids, but commercially available systems often do not enable res...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer International Publishing
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10020259/ https://www.ncbi.nlm.nih.gov/pubmed/36929461 http://dx.doi.org/10.1007/s00018-023-04748-1 |
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author | Tartagni, Ottavia Borók, Alexandra Mensà, Emanuela Bonyár, Attila Monti, Barbara Hofkens, Johan Porcelli, Anna Maria Zuccheri, Giampaolo |
author_facet | Tartagni, Ottavia Borók, Alexandra Mensà, Emanuela Bonyár, Attila Monti, Barbara Hofkens, Johan Porcelli, Anna Maria Zuccheri, Giampaolo |
author_sort | Tartagni, Ottavia |
collection | PubMed |
description | Multicellular tumor spheroids are rapidly emerging as an improved in vitro model with respect to more traditional 2D culturing. Microwell culturing is a simple and accessible method for generating a large number of uniformly sized spheroids, but commercially available systems often do not enable researchers to perform complete culturing and analysis pipelines and the mechanical properties of their culture environment are not commonly matching those of the target tissue. We herein report a simple method to obtain custom-designed self-built microwell arrays made of polydimethylsiloxane or agarose for uniform 3D cell structure generation. Such materials can provide an environment of tunable mechanical flexibility. We developed protocols to culture a variety of cancer and non-cancer cell lines in such devices and to perform molecular and imaging characterizations of the spheroid growth, viability, and response to pharmacological treatments. Hundreds of tumor spheroids grow (in scaffolded or scaffold-free conditions) at homogeneous rates and can be harvested at will. Microscopy imaging can be performed in situ during or at the end of the culture. Fluorescence (confocal) microscopy can be performed after in situ staining while retaining the geographic arrangement of spheroids in the plate wells. This platform can enable statistically robust investigations on cancer biology and screening of drug treatments. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00018-023-04748-1. |
format | Online Article Text |
id | pubmed-10020259 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Springer International Publishing |
record_format | MEDLINE/PubMed |
spelling | pubmed-100202592023-03-18 Microstructured soft devices for the growth and analysis of populations of homogenous multicellular tumor spheroids Tartagni, Ottavia Borók, Alexandra Mensà, Emanuela Bonyár, Attila Monti, Barbara Hofkens, Johan Porcelli, Anna Maria Zuccheri, Giampaolo Cell Mol Life Sci Original Article Multicellular tumor spheroids are rapidly emerging as an improved in vitro model with respect to more traditional 2D culturing. Microwell culturing is a simple and accessible method for generating a large number of uniformly sized spheroids, but commercially available systems often do not enable researchers to perform complete culturing and analysis pipelines and the mechanical properties of their culture environment are not commonly matching those of the target tissue. We herein report a simple method to obtain custom-designed self-built microwell arrays made of polydimethylsiloxane or agarose for uniform 3D cell structure generation. Such materials can provide an environment of tunable mechanical flexibility. We developed protocols to culture a variety of cancer and non-cancer cell lines in such devices and to perform molecular and imaging characterizations of the spheroid growth, viability, and response to pharmacological treatments. Hundreds of tumor spheroids grow (in scaffolded or scaffold-free conditions) at homogeneous rates and can be harvested at will. Microscopy imaging can be performed in situ during or at the end of the culture. Fluorescence (confocal) microscopy can be performed after in situ staining while retaining the geographic arrangement of spheroids in the plate wells. This platform can enable statistically robust investigations on cancer biology and screening of drug treatments. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00018-023-04748-1. Springer International Publishing 2023-03-16 2023 /pmc/articles/PMC10020259/ /pubmed/36929461 http://dx.doi.org/10.1007/s00018-023-04748-1 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Original Article Tartagni, Ottavia Borók, Alexandra Mensà, Emanuela Bonyár, Attila Monti, Barbara Hofkens, Johan Porcelli, Anna Maria Zuccheri, Giampaolo Microstructured soft devices for the growth and analysis of populations of homogenous multicellular tumor spheroids |
title | Microstructured soft devices for the growth and analysis of populations of homogenous multicellular tumor spheroids |
title_full | Microstructured soft devices for the growth and analysis of populations of homogenous multicellular tumor spheroids |
title_fullStr | Microstructured soft devices for the growth and analysis of populations of homogenous multicellular tumor spheroids |
title_full_unstemmed | Microstructured soft devices for the growth and analysis of populations of homogenous multicellular tumor spheroids |
title_short | Microstructured soft devices for the growth and analysis of populations of homogenous multicellular tumor spheroids |
title_sort | microstructured soft devices for the growth and analysis of populations of homogenous multicellular tumor spheroids |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10020259/ https://www.ncbi.nlm.nih.gov/pubmed/36929461 http://dx.doi.org/10.1007/s00018-023-04748-1 |
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