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Identification of extracellular matrix proteins secreted by human dermal fibroblasts cultured in 3D electrospun scaffolds

The appreciation that cell interactions in tissues is dependent on their three dimensional (3D) distribution has stimulated the development of 3D cell culture models. We constructed an artificial 3D tumour by culturing human breast cancer JIMT-1 cells and human dermal fibroblasts (HDFs) in a 3D netw...

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Autores principales: Malakpour-Permlid, Atena, Buzzi, Irina, Hegardt, Cecilia, Johansson, Fredrik, Oredsson, Stina
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7988018/
https://www.ncbi.nlm.nih.gov/pubmed/33758206
http://dx.doi.org/10.1038/s41598-021-85742-0
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author Malakpour-Permlid, Atena
Buzzi, Irina
Hegardt, Cecilia
Johansson, Fredrik
Oredsson, Stina
author_facet Malakpour-Permlid, Atena
Buzzi, Irina
Hegardt, Cecilia
Johansson, Fredrik
Oredsson, Stina
author_sort Malakpour-Permlid, Atena
collection PubMed
description The appreciation that cell interactions in tissues is dependent on their three dimensional (3D) distribution has stimulated the development of 3D cell culture models. We constructed an artificial 3D tumour by culturing human breast cancer JIMT-1 cells and human dermal fibroblasts (HDFs) in a 3D network of electrospun polycaprolactone fibres. Here, we investigate ECM components produced by the cells in the artificial 3D tumour, which is an important step in validating the model. Immunostaining and confocal fluorescence microscopy show that the ECM proteins fibronectin, collagen I, and laminin are deposited throughout the entire 3D structure. Secreted soluble factors including matrix metalloproteinases (MMPs) and interleukine-6 (IL-6) were analysed in collected medium and were found to be mainly derived from the HDFs. Treatment with transforming growth factor-β1 (TGF-β1), a major cytokine found in a tumour, significantly alters the MMP activity and IL-6 concentration. In addition, TGF-β1 treatment, changes the morphology of the HDFs to become more elongated and with increased linearized actin filaments compared to non-treated HDFs. Collectively, these novel findings suggest that the artificial 3D tumour displays a clear cell distribution and ECM deposition that resembles a tumour environment in vivo, suggesting an innovative biological model to study a human tumour.
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spelling pubmed-79880182021-03-25 Identification of extracellular matrix proteins secreted by human dermal fibroblasts cultured in 3D electrospun scaffolds Malakpour-Permlid, Atena Buzzi, Irina Hegardt, Cecilia Johansson, Fredrik Oredsson, Stina Sci Rep Article The appreciation that cell interactions in tissues is dependent on their three dimensional (3D) distribution has stimulated the development of 3D cell culture models. We constructed an artificial 3D tumour by culturing human breast cancer JIMT-1 cells and human dermal fibroblasts (HDFs) in a 3D network of electrospun polycaprolactone fibres. Here, we investigate ECM components produced by the cells in the artificial 3D tumour, which is an important step in validating the model. Immunostaining and confocal fluorescence microscopy show that the ECM proteins fibronectin, collagen I, and laminin are deposited throughout the entire 3D structure. Secreted soluble factors including matrix metalloproteinases (MMPs) and interleukine-6 (IL-6) were analysed in collected medium and were found to be mainly derived from the HDFs. Treatment with transforming growth factor-β1 (TGF-β1), a major cytokine found in a tumour, significantly alters the MMP activity and IL-6 concentration. In addition, TGF-β1 treatment, changes the morphology of the HDFs to become more elongated and with increased linearized actin filaments compared to non-treated HDFs. Collectively, these novel findings suggest that the artificial 3D tumour displays a clear cell distribution and ECM deposition that resembles a tumour environment in vivo, suggesting an innovative biological model to study a human tumour. Nature Publishing Group UK 2021-03-23 /pmc/articles/PMC7988018/ /pubmed/33758206 http://dx.doi.org/10.1038/s41598-021-85742-0 Text en © The Author(s) 2021 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Malakpour-Permlid, Atena
Buzzi, Irina
Hegardt, Cecilia
Johansson, Fredrik
Oredsson, Stina
Identification of extracellular matrix proteins secreted by human dermal fibroblasts cultured in 3D electrospun scaffolds
title Identification of extracellular matrix proteins secreted by human dermal fibroblasts cultured in 3D electrospun scaffolds
title_full Identification of extracellular matrix proteins secreted by human dermal fibroblasts cultured in 3D electrospun scaffolds
title_fullStr Identification of extracellular matrix proteins secreted by human dermal fibroblasts cultured in 3D electrospun scaffolds
title_full_unstemmed Identification of extracellular matrix proteins secreted by human dermal fibroblasts cultured in 3D electrospun scaffolds
title_short Identification of extracellular matrix proteins secreted by human dermal fibroblasts cultured in 3D electrospun scaffolds
title_sort identification of extracellular matrix proteins secreted by human dermal fibroblasts cultured in 3d electrospun scaffolds
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7988018/
https://www.ncbi.nlm.nih.gov/pubmed/33758206
http://dx.doi.org/10.1038/s41598-021-85742-0
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