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A tunable physiomimetic stretch system evaluated with precision cut lung slices and recellularized human lung scaffolds
Breathing exposes lung cells to continual mechanical stimuli, which is part of the microenvironmental signals directing cellular functions together with the extracellular matrix (ECM). Therefore, developing systems that incorporate both stimuli is urgent to fully understand cell behavior. This study...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9574011/ https://www.ncbi.nlm.nih.gov/pubmed/36263353 http://dx.doi.org/10.3389/fbioe.2022.995460 |
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author | Rosmark, Oskar Ibáñez-Fonseca, Arturo Thorsson, Johan Dellgren, Göran Hallgren, Oskar Larsson Callerfelt, Anna-Karin Elowsson, Linda Westergren-Thorsson, Gunilla |
author_facet | Rosmark, Oskar Ibáñez-Fonseca, Arturo Thorsson, Johan Dellgren, Göran Hallgren, Oskar Larsson Callerfelt, Anna-Karin Elowsson, Linda Westergren-Thorsson, Gunilla |
author_sort | Rosmark, Oskar |
collection | PubMed |
description | Breathing exposes lung cells to continual mechanical stimuli, which is part of the microenvironmental signals directing cellular functions together with the extracellular matrix (ECM). Therefore, developing systems that incorporate both stimuli is urgent to fully understand cell behavior. This study aims to introduce a novel in vitro culture methodology combining a cyclic stretch that simulates in vivo breathing with 3D cell culture platforms in the form of decellularized lung slices (DLS) and precision cut lung slices (PCLS). To this end, we have constructed a device that mimics the amplitudes and frequencies of distensions seen in the breathing human lung. For its validation, we cultured H441 lung epithelial cells in human DLS exposed to 16 stretch cycles per minute with a 10% stretch amplitude. Cell viability (resazurin reduction), proliferation (Ki-67) and YAP1 activation were evaluated at 24 and 96 h by immunohistochemistry, while the expression of SFTPB, COL3A1, COL4A3 and LAMA5 was evaluated by qPCR. Cyclic stretch induced an increase in SFTPB expression after 24 h without a concomitant increase in the stretch responsive gene YAP1. Moreover, the ECM milieu lowered the expression of the basement membrane protein genes COL4A3 and LAMA5 compared to tissue culture plastic control cultures, but no effect was observed by the mechanical stimuli. The device also confirmed good compatibility with PCLS culture, showing preserved morphology and metabolism in rat PCLS after 72 h of mechanical stretch. Thus, we present a novel device and methodology for the easy assembling and study of lung tissue slice cultures subjected to physiomimetic mechanical stimuli, which shows promise for future studies of cell and tissue function in a lung ECM milieu with physiological or pathological mechanical stimuli. |
format | Online Article Text |
id | pubmed-9574011 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-95740112022-10-18 A tunable physiomimetic stretch system evaluated with precision cut lung slices and recellularized human lung scaffolds Rosmark, Oskar Ibáñez-Fonseca, Arturo Thorsson, Johan Dellgren, Göran Hallgren, Oskar Larsson Callerfelt, Anna-Karin Elowsson, Linda Westergren-Thorsson, Gunilla Front Bioeng Biotechnol Bioengineering and Biotechnology Breathing exposes lung cells to continual mechanical stimuli, which is part of the microenvironmental signals directing cellular functions together with the extracellular matrix (ECM). Therefore, developing systems that incorporate both stimuli is urgent to fully understand cell behavior. This study aims to introduce a novel in vitro culture methodology combining a cyclic stretch that simulates in vivo breathing with 3D cell culture platforms in the form of decellularized lung slices (DLS) and precision cut lung slices (PCLS). To this end, we have constructed a device that mimics the amplitudes and frequencies of distensions seen in the breathing human lung. For its validation, we cultured H441 lung epithelial cells in human DLS exposed to 16 stretch cycles per minute with a 10% stretch amplitude. Cell viability (resazurin reduction), proliferation (Ki-67) and YAP1 activation were evaluated at 24 and 96 h by immunohistochemistry, while the expression of SFTPB, COL3A1, COL4A3 and LAMA5 was evaluated by qPCR. Cyclic stretch induced an increase in SFTPB expression after 24 h without a concomitant increase in the stretch responsive gene YAP1. Moreover, the ECM milieu lowered the expression of the basement membrane protein genes COL4A3 and LAMA5 compared to tissue culture plastic control cultures, but no effect was observed by the mechanical stimuli. The device also confirmed good compatibility with PCLS culture, showing preserved morphology and metabolism in rat PCLS after 72 h of mechanical stretch. Thus, we present a novel device and methodology for the easy assembling and study of lung tissue slice cultures subjected to physiomimetic mechanical stimuli, which shows promise for future studies of cell and tissue function in a lung ECM milieu with physiological or pathological mechanical stimuli. Frontiers Media S.A. 2022-10-03 /pmc/articles/PMC9574011/ /pubmed/36263353 http://dx.doi.org/10.3389/fbioe.2022.995460 Text en Copyright © 2022 Rosmark, Ibáñez-Fonseca, Thorsson, Dellgren, Hallgren, Larsson Callerfelt, Elowsson and Westergren-Thorsson. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Bioengineering and Biotechnology Rosmark, Oskar Ibáñez-Fonseca, Arturo Thorsson, Johan Dellgren, Göran Hallgren, Oskar Larsson Callerfelt, Anna-Karin Elowsson, Linda Westergren-Thorsson, Gunilla A tunable physiomimetic stretch system evaluated with precision cut lung slices and recellularized human lung scaffolds |
title | A tunable physiomimetic stretch system evaluated with precision cut lung slices and recellularized human lung scaffolds |
title_full | A tunable physiomimetic stretch system evaluated with precision cut lung slices and recellularized human lung scaffolds |
title_fullStr | A tunable physiomimetic stretch system evaluated with precision cut lung slices and recellularized human lung scaffolds |
title_full_unstemmed | A tunable physiomimetic stretch system evaluated with precision cut lung slices and recellularized human lung scaffolds |
title_short | A tunable physiomimetic stretch system evaluated with precision cut lung slices and recellularized human lung scaffolds |
title_sort | tunable physiomimetic stretch system evaluated with precision cut lung slices and recellularized human lung scaffolds |
topic | Bioengineering and Biotechnology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9574011/ https://www.ncbi.nlm.nih.gov/pubmed/36263353 http://dx.doi.org/10.3389/fbioe.2022.995460 |
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