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Fibronectin localization and fibrillization are affected by the presence of serum in culture media

In vitro models of fibrotic phenomena are often based on the fibroblast-myofibroblast transition as the contraction-triggering cellular event. There are, however, multiple sources of concern regarding the appropriateness of such models; a first and widely investigated issue is the often inappropriat...

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Autores principales: Siani, Alessandro, Khaw, Rong R., Manley, Oliver W. G., Tirella, Annalisa, Cellesi, Francesco, Donno, Roberto, Tirelli, Nicola
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4369722/
https://www.ncbi.nlm.nih.gov/pubmed/25797118
http://dx.doi.org/10.1038/srep09278
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author Siani, Alessandro
Khaw, Rong R.
Manley, Oliver W. G.
Tirella, Annalisa
Cellesi, Francesco
Donno, Roberto
Tirelli, Nicola
author_facet Siani, Alessandro
Khaw, Rong R.
Manley, Oliver W. G.
Tirella, Annalisa
Cellesi, Francesco
Donno, Roberto
Tirelli, Nicola
author_sort Siani, Alessandro
collection PubMed
description In vitro models of fibrotic phenomena are often based on the fibroblast-myofibroblast transition as the contraction-triggering cellular event. There are, however, multiple sources of concern regarding the appropriateness of such models; a first and widely investigated issue is the often inappropriate nature of the interactions between mesenchymal cells and surrounding/underlying matrix/substrate. A second set of problems concerns the composition of the fluid phase, which includes both dispersed/dissolved paracrine messengers and matrix elements. In this study, we have focused on the effects that serum may generate. We have observed that A) serum causes high variability in the expression of typical markers of myofibroblast differentiation (ED-A fibronectin and α-Smooth Muscle Actin) upon treatment with TGF-β1; this is probably due to intrinsic variability of cytokine concentrations in different batches of serum. B) the fibrillization of endogenous fibronectin is partially hampered and its localization changed from ventral (on the substrate) to dorsal (upper surface); the latter morphology appears to be largely overlooked in literature, even though it may have a significant role in terms of mechanotransductive signaling. This quite dramatic change possibly occurs as a result of competition with serum proteins, although our data seem to rule out a direct role of serum fibronectin.
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spelling pubmed-43697222015-04-06 Fibronectin localization and fibrillization are affected by the presence of serum in culture media Siani, Alessandro Khaw, Rong R. Manley, Oliver W. G. Tirella, Annalisa Cellesi, Francesco Donno, Roberto Tirelli, Nicola Sci Rep Article In vitro models of fibrotic phenomena are often based on the fibroblast-myofibroblast transition as the contraction-triggering cellular event. There are, however, multiple sources of concern regarding the appropriateness of such models; a first and widely investigated issue is the often inappropriate nature of the interactions between mesenchymal cells and surrounding/underlying matrix/substrate. A second set of problems concerns the composition of the fluid phase, which includes both dispersed/dissolved paracrine messengers and matrix elements. In this study, we have focused on the effects that serum may generate. We have observed that A) serum causes high variability in the expression of typical markers of myofibroblast differentiation (ED-A fibronectin and α-Smooth Muscle Actin) upon treatment with TGF-β1; this is probably due to intrinsic variability of cytokine concentrations in different batches of serum. B) the fibrillization of endogenous fibronectin is partially hampered and its localization changed from ventral (on the substrate) to dorsal (upper surface); the latter morphology appears to be largely overlooked in literature, even though it may have a significant role in terms of mechanotransductive signaling. This quite dramatic change possibly occurs as a result of competition with serum proteins, although our data seem to rule out a direct role of serum fibronectin. Nature Publishing Group 2015-03-23 /pmc/articles/PMC4369722/ /pubmed/25797118 http://dx.doi.org/10.1038/srep09278 Text en Copyright © 2015, Macmillan Publishers Limited. All rights reserved http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder in order to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Siani, Alessandro
Khaw, Rong R.
Manley, Oliver W. G.
Tirella, Annalisa
Cellesi, Francesco
Donno, Roberto
Tirelli, Nicola
Fibronectin localization and fibrillization are affected by the presence of serum in culture media
title Fibronectin localization and fibrillization are affected by the presence of serum in culture media
title_full Fibronectin localization and fibrillization are affected by the presence of serum in culture media
title_fullStr Fibronectin localization and fibrillization are affected by the presence of serum in culture media
title_full_unstemmed Fibronectin localization and fibrillization are affected by the presence of serum in culture media
title_short Fibronectin localization and fibrillization are affected by the presence of serum in culture media
title_sort fibronectin localization and fibrillization are affected by the presence of serum in culture media
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4369722/
https://www.ncbi.nlm.nih.gov/pubmed/25797118
http://dx.doi.org/10.1038/srep09278
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