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Electron microscopy imaging and mechanical characterization of T47D multicellular tumor spheroids–Older spheroids reduce interstitial space and become stiffer

Multicellular cancer spheroids are an in vitro tissue model that mimics the three-dimensional microenvironment. As spheroids grow, they develop the gradients of oxygen, nutrients, and catabolites, affecting crucial tumor characteristics such as proliferation and treatment responses. The measurement...

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Autores principales: Madhavan, Mathangi, Jaiswal, Devina, Karlberg, Sarah, Duggan, Alexis, Almarshad, Hassan A., Claffey, Kevin P., Hoshino, Kazunori
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10212087/
https://www.ncbi.nlm.nih.gov/pubmed/37228139
http://dx.doi.org/10.1371/journal.pone.0286291
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author Madhavan, Mathangi
Jaiswal, Devina
Karlberg, Sarah
Duggan, Alexis
Almarshad, Hassan A.
Claffey, Kevin P.
Hoshino, Kazunori
author_facet Madhavan, Mathangi
Jaiswal, Devina
Karlberg, Sarah
Duggan, Alexis
Almarshad, Hassan A.
Claffey, Kevin P.
Hoshino, Kazunori
author_sort Madhavan, Mathangi
collection PubMed
description Multicellular cancer spheroids are an in vitro tissue model that mimics the three-dimensional microenvironment. As spheroids grow, they develop the gradients of oxygen, nutrients, and catabolites, affecting crucial tumor characteristics such as proliferation and treatment responses. The measurement of spheroid stiffness provides a quantitative measure to evaluate such structural changes over time. In this report, we measured the stiffness of size-matched day 5 and day 20 tumor spheroids using a custom-built microscale force sensor and conducted transmission electron microscopy (TEM) imaging to compare the internal structures. We found that older spheroids reduce interstitial spaces in the core region and became significantly stiffer. The measured elastic moduli were 260±100 and 680±150 Pa, for day 5 and day 20 spheroids, respectively. The day 20 spheroids showed an optically dark region in the center. Analyzing the high-resolution TEM images of spheroid middle sections across the diameter showed that the cells in the inner region of the day 20 spheroids are significantly larger and more closely packed than those in the outer regions. On the other hand, the day 5 spheroids did not show a significant difference between the inner and outer regions. The observed reduction of the interstitial space may be one factor that contributes to stiffer older spheroids.
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spelling pubmed-102120872023-05-26 Electron microscopy imaging and mechanical characterization of T47D multicellular tumor spheroids–Older spheroids reduce interstitial space and become stiffer Madhavan, Mathangi Jaiswal, Devina Karlberg, Sarah Duggan, Alexis Almarshad, Hassan A. Claffey, Kevin P. Hoshino, Kazunori PLoS One Research Article Multicellular cancer spheroids are an in vitro tissue model that mimics the three-dimensional microenvironment. As spheroids grow, they develop the gradients of oxygen, nutrients, and catabolites, affecting crucial tumor characteristics such as proliferation and treatment responses. The measurement of spheroid stiffness provides a quantitative measure to evaluate such structural changes over time. In this report, we measured the stiffness of size-matched day 5 and day 20 tumor spheroids using a custom-built microscale force sensor and conducted transmission electron microscopy (TEM) imaging to compare the internal structures. We found that older spheroids reduce interstitial spaces in the core region and became significantly stiffer. The measured elastic moduli were 260±100 and 680±150 Pa, for day 5 and day 20 spheroids, respectively. The day 20 spheroids showed an optically dark region in the center. Analyzing the high-resolution TEM images of spheroid middle sections across the diameter showed that the cells in the inner region of the day 20 spheroids are significantly larger and more closely packed than those in the outer regions. On the other hand, the day 5 spheroids did not show a significant difference between the inner and outer regions. The observed reduction of the interstitial space may be one factor that contributes to stiffer older spheroids. Public Library of Science 2023-05-25 /pmc/articles/PMC10212087/ /pubmed/37228139 http://dx.doi.org/10.1371/journal.pone.0286291 Text en © 2023 Madhavan et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Madhavan, Mathangi
Jaiswal, Devina
Karlberg, Sarah
Duggan, Alexis
Almarshad, Hassan A.
Claffey, Kevin P.
Hoshino, Kazunori
Electron microscopy imaging and mechanical characterization of T47D multicellular tumor spheroids–Older spheroids reduce interstitial space and become stiffer
title Electron microscopy imaging and mechanical characterization of T47D multicellular tumor spheroids–Older spheroids reduce interstitial space and become stiffer
title_full Electron microscopy imaging and mechanical characterization of T47D multicellular tumor spheroids–Older spheroids reduce interstitial space and become stiffer
title_fullStr Electron microscopy imaging and mechanical characterization of T47D multicellular tumor spheroids–Older spheroids reduce interstitial space and become stiffer
title_full_unstemmed Electron microscopy imaging and mechanical characterization of T47D multicellular tumor spheroids–Older spheroids reduce interstitial space and become stiffer
title_short Electron microscopy imaging and mechanical characterization of T47D multicellular tumor spheroids–Older spheroids reduce interstitial space and become stiffer
title_sort electron microscopy imaging and mechanical characterization of t47d multicellular tumor spheroids–older spheroids reduce interstitial space and become stiffer
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10212087/
https://www.ncbi.nlm.nih.gov/pubmed/37228139
http://dx.doi.org/10.1371/journal.pone.0286291
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