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The Impact of Experimental Conditions on Cell Mechanics as Measured with Nanoindentation

The evaluation of cell elasticity is becoming increasingly significant, since it is now known that it impacts physiological mechanisms, such as stem cell differentiation and embryogenesis, as well as pathological processes, such as cancer invasiveness and endothelial senescence. However, the results...

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Autores principales: Zambito, Martina, Viti, Federica, Bosio, Alessia G., Ceccherini, Isabella, Florio, Tullio, Vassalli, Massimo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10097320/
https://www.ncbi.nlm.nih.gov/pubmed/37049284
http://dx.doi.org/10.3390/nano13071190
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author Zambito, Martina
Viti, Federica
Bosio, Alessia G.
Ceccherini, Isabella
Florio, Tullio
Vassalli, Massimo
author_facet Zambito, Martina
Viti, Federica
Bosio, Alessia G.
Ceccherini, Isabella
Florio, Tullio
Vassalli, Massimo
author_sort Zambito, Martina
collection PubMed
description The evaluation of cell elasticity is becoming increasingly significant, since it is now known that it impacts physiological mechanisms, such as stem cell differentiation and embryogenesis, as well as pathological processes, such as cancer invasiveness and endothelial senescence. However, the results of single-cell mechanical measurements vary considerably, not only due to systematic instrumental errors but also due to the dynamic and non-homogenous nature of the sample. In this work, relying on Chiaro nanoindenter (Optics11Life), we characterized in depth the nanoindentation experimental procedure, in order to highlight whether and how experimental conditions could affect measurements of living cell stiffness. We demonstrated that the procedure can be quite insensitive to technical replicates and that several biological conditions, such as cell confluency, starvation and passage, significantly impact the results. Experiments should be designed to maximally avoid inhomogeneous scenarios to avoid divergences in the measured phenotype.
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spelling pubmed-100973202023-04-13 The Impact of Experimental Conditions on Cell Mechanics as Measured with Nanoindentation Zambito, Martina Viti, Federica Bosio, Alessia G. Ceccherini, Isabella Florio, Tullio Vassalli, Massimo Nanomaterials (Basel) Article The evaluation of cell elasticity is becoming increasingly significant, since it is now known that it impacts physiological mechanisms, such as stem cell differentiation and embryogenesis, as well as pathological processes, such as cancer invasiveness and endothelial senescence. However, the results of single-cell mechanical measurements vary considerably, not only due to systematic instrumental errors but also due to the dynamic and non-homogenous nature of the sample. In this work, relying on Chiaro nanoindenter (Optics11Life), we characterized in depth the nanoindentation experimental procedure, in order to highlight whether and how experimental conditions could affect measurements of living cell stiffness. We demonstrated that the procedure can be quite insensitive to technical replicates and that several biological conditions, such as cell confluency, starvation and passage, significantly impact the results. Experiments should be designed to maximally avoid inhomogeneous scenarios to avoid divergences in the measured phenotype. MDPI 2023-03-27 /pmc/articles/PMC10097320/ /pubmed/37049284 http://dx.doi.org/10.3390/nano13071190 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Zambito, Martina
Viti, Federica
Bosio, Alessia G.
Ceccherini, Isabella
Florio, Tullio
Vassalli, Massimo
The Impact of Experimental Conditions on Cell Mechanics as Measured with Nanoindentation
title The Impact of Experimental Conditions on Cell Mechanics as Measured with Nanoindentation
title_full The Impact of Experimental Conditions on Cell Mechanics as Measured with Nanoindentation
title_fullStr The Impact of Experimental Conditions on Cell Mechanics as Measured with Nanoindentation
title_full_unstemmed The Impact of Experimental Conditions on Cell Mechanics as Measured with Nanoindentation
title_short The Impact of Experimental Conditions on Cell Mechanics as Measured with Nanoindentation
title_sort impact of experimental conditions on cell mechanics as measured with nanoindentation
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10097320/
https://www.ncbi.nlm.nih.gov/pubmed/37049284
http://dx.doi.org/10.3390/nano13071190
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