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A new method for the study of biophysical and morphological parameters in 3D cell cultures: Evaluation in LoVo spheroids treated with crizotinib

Three-dimensional (3D) culture systems like tumor spheroids represent useful in vitro models for drug screening and more broadly for cancer biology research, but the generation of uniform populations of spheroids remains challenging. The possibility to properly characterize spheroid properties would...

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Autores principales: Sargenti, Azzurra, Musmeci, Francesco, Cavallo, Carola, Mazzeschi, Martina, Bonetti, Simone, Pasqua, Simone, Bacchi, Francesco, Filardo, Giuseppe, Gazzola, Daniele, Lauriola, Mattia, Santi, Spartaco
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8186796/
https://www.ncbi.nlm.nih.gov/pubmed/34101765
http://dx.doi.org/10.1371/journal.pone.0252907
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author Sargenti, Azzurra
Musmeci, Francesco
Cavallo, Carola
Mazzeschi, Martina
Bonetti, Simone
Pasqua, Simone
Bacchi, Francesco
Filardo, Giuseppe
Gazzola, Daniele
Lauriola, Mattia
Santi, Spartaco
author_facet Sargenti, Azzurra
Musmeci, Francesco
Cavallo, Carola
Mazzeschi, Martina
Bonetti, Simone
Pasqua, Simone
Bacchi, Francesco
Filardo, Giuseppe
Gazzola, Daniele
Lauriola, Mattia
Santi, Spartaco
author_sort Sargenti, Azzurra
collection PubMed
description Three-dimensional (3D) culture systems like tumor spheroids represent useful in vitro models for drug screening and more broadly for cancer biology research, but the generation of uniform populations of spheroids remains challenging. The possibility to properly characterize spheroid properties would increase the reliability of these models. To address this issue different analysis were combined: i) a new device and relative analytical method for the accurate, simultaneous, and rapid measurement of mass density, weight, and size of spheroids, ii) confocal imaging, and iii) protein quantification, in a clinically relevant 3D model. The LoVo colon cancer cell line forming spheroids, treated with crizotinib (CZB) an ATP-competitive small-molecule inhibitor of the receptor tyrosine kinases, was employed to study and assess the correlation between biophysical and morphological parameters in both live and fixed cells. The new fluidic-based measurements allowed a robust phenotypical characterization of the spheroids structure, offering insights on the spheroids bulk and an accurate measurement of the tumor density. This analysis helps overcome the technical limits of the imaging that hardly penetrates the thickness of 3D structures. Accordingly, we were able to document that CZB treatment has an impact on mass density, which represents a key marker characterizing cancer cell treatment. Spheroid culture is the ultimate technology in drug discovery and the adoption of such precise measurement of the tumor characteristics can represent a key step forward for the accurate testing of treatment’s potential in 3D in vitro models.
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spelling pubmed-81867962021-06-16 A new method for the study of biophysical and morphological parameters in 3D cell cultures: Evaluation in LoVo spheroids treated with crizotinib Sargenti, Azzurra Musmeci, Francesco Cavallo, Carola Mazzeschi, Martina Bonetti, Simone Pasqua, Simone Bacchi, Francesco Filardo, Giuseppe Gazzola, Daniele Lauriola, Mattia Santi, Spartaco PLoS One Research Article Three-dimensional (3D) culture systems like tumor spheroids represent useful in vitro models for drug screening and more broadly for cancer biology research, but the generation of uniform populations of spheroids remains challenging. The possibility to properly characterize spheroid properties would increase the reliability of these models. To address this issue different analysis were combined: i) a new device and relative analytical method for the accurate, simultaneous, and rapid measurement of mass density, weight, and size of spheroids, ii) confocal imaging, and iii) protein quantification, in a clinically relevant 3D model. The LoVo colon cancer cell line forming spheroids, treated with crizotinib (CZB) an ATP-competitive small-molecule inhibitor of the receptor tyrosine kinases, was employed to study and assess the correlation between biophysical and morphological parameters in both live and fixed cells. The new fluidic-based measurements allowed a robust phenotypical characterization of the spheroids structure, offering insights on the spheroids bulk and an accurate measurement of the tumor density. This analysis helps overcome the technical limits of the imaging that hardly penetrates the thickness of 3D structures. Accordingly, we were able to document that CZB treatment has an impact on mass density, which represents a key marker characterizing cancer cell treatment. Spheroid culture is the ultimate technology in drug discovery and the adoption of such precise measurement of the tumor characteristics can represent a key step forward for the accurate testing of treatment’s potential in 3D in vitro models. Public Library of Science 2021-06-08 /pmc/articles/PMC8186796/ /pubmed/34101765 http://dx.doi.org/10.1371/journal.pone.0252907 Text en © 2021 Sargenti 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
Sargenti, Azzurra
Musmeci, Francesco
Cavallo, Carola
Mazzeschi, Martina
Bonetti, Simone
Pasqua, Simone
Bacchi, Francesco
Filardo, Giuseppe
Gazzola, Daniele
Lauriola, Mattia
Santi, Spartaco
A new method for the study of biophysical and morphological parameters in 3D cell cultures: Evaluation in LoVo spheroids treated with crizotinib
title A new method for the study of biophysical and morphological parameters in 3D cell cultures: Evaluation in LoVo spheroids treated with crizotinib
title_full A new method for the study of biophysical and morphological parameters in 3D cell cultures: Evaluation in LoVo spheroids treated with crizotinib
title_fullStr A new method for the study of biophysical and morphological parameters in 3D cell cultures: Evaluation in LoVo spheroids treated with crizotinib
title_full_unstemmed A new method for the study of biophysical and morphological parameters in 3D cell cultures: Evaluation in LoVo spheroids treated with crizotinib
title_short A new method for the study of biophysical and morphological parameters in 3D cell cultures: Evaluation in LoVo spheroids treated with crizotinib
title_sort new method for the study of biophysical and morphological parameters in 3d cell cultures: evaluation in lovo spheroids treated with crizotinib
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8186796/
https://www.ncbi.nlm.nih.gov/pubmed/34101765
http://dx.doi.org/10.1371/journal.pone.0252907
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