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Cell Culture Platforms with Controllable Stiffness for Chick Embryonic Cardiomyocytes

For several years, cell culture techniques have been physiologically relevant to understand living organisms both structurally and functionally, aiming at preserving as carefully as possible the in vivo integrity and function of the cells. However, when studying cardiac cells, glass or plastic Petri...

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Autores principales: Durán-Pastén, María Luisa, Cortes, Daniela, Valencia-Amaya, Alan E., King, Santiago, González-Gómez, Gertrudis Hortensia, Hautefeuille, Mathieu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6630216/
https://www.ncbi.nlm.nih.gov/pubmed/31105218
http://dx.doi.org/10.3390/biomimetics4020033
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author Durán-Pastén, María Luisa
Cortes, Daniela
Valencia-Amaya, Alan E.
King, Santiago
González-Gómez, Gertrudis Hortensia
Hautefeuille, Mathieu
author_facet Durán-Pastén, María Luisa
Cortes, Daniela
Valencia-Amaya, Alan E.
King, Santiago
González-Gómez, Gertrudis Hortensia
Hautefeuille, Mathieu
author_sort Durán-Pastén, María Luisa
collection PubMed
description For several years, cell culture techniques have been physiologically relevant to understand living organisms both structurally and functionally, aiming at preserving as carefully as possible the in vivo integrity and function of the cells. However, when studying cardiac cells, glass or plastic Petri dishes and culture-coated plates lack important cues that do not allow to maintain the desired phenotype, especially for primary cell culture. In this work, we show that microscaffolds made with polydimethylsiloxane (PDMS) enable modulating the stiffness of the surface of the culture substrate and this originates different patterns of adhesion, self-organization, and synchronized or propagated activity in the culture of chick embryonic cardiomyocytes. Thanks to the calcium imaging technique, we found that the substrate stiffness affected cardiomyocyte adhesion, as well as the calcium signal propagation in the formed tissue. The patterns of activity shown by the calcium fluorescence variations are reliable clues of the functional organization achieved by the cell layers. We found that PDMS substrates with a stiffness of 25 kPa did not allow the formation of cell layers and therefore the optimal propagation of the intracellular calcium signals, while softer PDMS substrates with Young’s modulus within the physiological in vivo reported range did permit synchronized and coordinated contractility and intracellular calcium activity. This type of methodology allows us to study phenomena such as arrhythmias. For example, the occurrence of synchronized activity or rotors that can initiate or maintain cardiac arrhythmias can be reproduced on different substrates for study, so that replacement tissues or patches can be better designed.
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spelling pubmed-66302162019-08-19 Cell Culture Platforms with Controllable Stiffness for Chick Embryonic Cardiomyocytes Durán-Pastén, María Luisa Cortes, Daniela Valencia-Amaya, Alan E. King, Santiago González-Gómez, Gertrudis Hortensia Hautefeuille, Mathieu Biomimetics (Basel) Article For several years, cell culture techniques have been physiologically relevant to understand living organisms both structurally and functionally, aiming at preserving as carefully as possible the in vivo integrity and function of the cells. However, when studying cardiac cells, glass or plastic Petri dishes and culture-coated plates lack important cues that do not allow to maintain the desired phenotype, especially for primary cell culture. In this work, we show that microscaffolds made with polydimethylsiloxane (PDMS) enable modulating the stiffness of the surface of the culture substrate and this originates different patterns of adhesion, self-organization, and synchronized or propagated activity in the culture of chick embryonic cardiomyocytes. Thanks to the calcium imaging technique, we found that the substrate stiffness affected cardiomyocyte adhesion, as well as the calcium signal propagation in the formed tissue. The patterns of activity shown by the calcium fluorescence variations are reliable clues of the functional organization achieved by the cell layers. We found that PDMS substrates with a stiffness of 25 kPa did not allow the formation of cell layers and therefore the optimal propagation of the intracellular calcium signals, while softer PDMS substrates with Young’s modulus within the physiological in vivo reported range did permit synchronized and coordinated contractility and intracellular calcium activity. This type of methodology allows us to study phenomena such as arrhythmias. For example, the occurrence of synchronized activity or rotors that can initiate or maintain cardiac arrhythmias can be reproduced on different substrates for study, so that replacement tissues or patches can be better designed. MDPI 2019-04-27 /pmc/articles/PMC6630216/ /pubmed/31105218 http://dx.doi.org/10.3390/biomimetics4020033 Text en © 2019 by the authors. 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 (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Durán-Pastén, María Luisa
Cortes, Daniela
Valencia-Amaya, Alan E.
King, Santiago
González-Gómez, Gertrudis Hortensia
Hautefeuille, Mathieu
Cell Culture Platforms with Controllable Stiffness for Chick Embryonic Cardiomyocytes
title Cell Culture Platforms with Controllable Stiffness for Chick Embryonic Cardiomyocytes
title_full Cell Culture Platforms with Controllable Stiffness for Chick Embryonic Cardiomyocytes
title_fullStr Cell Culture Platforms with Controllable Stiffness for Chick Embryonic Cardiomyocytes
title_full_unstemmed Cell Culture Platforms with Controllable Stiffness for Chick Embryonic Cardiomyocytes
title_short Cell Culture Platforms with Controllable Stiffness for Chick Embryonic Cardiomyocytes
title_sort cell culture platforms with controllable stiffness for chick embryonic cardiomyocytes
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6630216/
https://www.ncbi.nlm.nih.gov/pubmed/31105218
http://dx.doi.org/10.3390/biomimetics4020033
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