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Epithelial restitution in 3D - Revealing biomechanical and physiochemical dynamics in intestinal organoids via fs laser nanosurgery
Intestinal organoids represent a three-dimensional cell culture system mimicking the mammalian intestine. The application of single-cell ablation for defined wounding via a femtosecond laser system within the crypt base allowed us to study cell dynamics during epithelial restitution. Neighboring cel...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10585398/ https://www.ncbi.nlm.nih.gov/pubmed/37867948 http://dx.doi.org/10.1016/j.isci.2023.108139 |
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author | Donath, Sören Seidler, Anna Elisabeth Mundin, Karlina Wenzel, Johannes Scholz, Jonas Gentemann, Lara Kalies, Julia Faix, Jan Ngezahayo, Anaclet Bleich, André Heisterkamp, Alexander Buettner, Manuela Kalies, Stefan |
author_facet | Donath, Sören Seidler, Anna Elisabeth Mundin, Karlina Wenzel, Johannes Scholz, Jonas Gentemann, Lara Kalies, Julia Faix, Jan Ngezahayo, Anaclet Bleich, André Heisterkamp, Alexander Buettner, Manuela Kalies, Stefan |
author_sort | Donath, Sören |
collection | PubMed |
description | Intestinal organoids represent a three-dimensional cell culture system mimicking the mammalian intestine. The application of single-cell ablation for defined wounding via a femtosecond laser system within the crypt base allowed us to study cell dynamics during epithelial restitution. Neighboring cells formed a contractile actin ring encircling the damaged cell, changed the cellular aspect ratio, and immediately closed the barrier. Using traction force microscopy, we observed major forces at the ablation site and additional forces on the crypt sides. Inhibitors of the actomyosin-based mobility of the cells led to the failure of restoring the barrier. Close to the ablation site, high-frequency calcium flickering and propagation of calcium waves occured that synchronized with the contraction of the epithelial layer. We observed an increased signal and nuclear translocation of YAP-1. In conclusion, our approach enabled, for the first time, to unveil the intricacies of epithelial restitution beyond in vivo models by employing precise laser-induced damage in colonoids. |
format | Online Article Text |
id | pubmed-10585398 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-105853982023-10-20 Epithelial restitution in 3D - Revealing biomechanical and physiochemical dynamics in intestinal organoids via fs laser nanosurgery Donath, Sören Seidler, Anna Elisabeth Mundin, Karlina Wenzel, Johannes Scholz, Jonas Gentemann, Lara Kalies, Julia Faix, Jan Ngezahayo, Anaclet Bleich, André Heisterkamp, Alexander Buettner, Manuela Kalies, Stefan iScience Article Intestinal organoids represent a three-dimensional cell culture system mimicking the mammalian intestine. The application of single-cell ablation for defined wounding via a femtosecond laser system within the crypt base allowed us to study cell dynamics during epithelial restitution. Neighboring cells formed a contractile actin ring encircling the damaged cell, changed the cellular aspect ratio, and immediately closed the barrier. Using traction force microscopy, we observed major forces at the ablation site and additional forces on the crypt sides. Inhibitors of the actomyosin-based mobility of the cells led to the failure of restoring the barrier. Close to the ablation site, high-frequency calcium flickering and propagation of calcium waves occured that synchronized with the contraction of the epithelial layer. We observed an increased signal and nuclear translocation of YAP-1. In conclusion, our approach enabled, for the first time, to unveil the intricacies of epithelial restitution beyond in vivo models by employing precise laser-induced damage in colonoids. Elsevier 2023-10-05 /pmc/articles/PMC10585398/ /pubmed/37867948 http://dx.doi.org/10.1016/j.isci.2023.108139 Text en © 2023 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Article Donath, Sören Seidler, Anna Elisabeth Mundin, Karlina Wenzel, Johannes Scholz, Jonas Gentemann, Lara Kalies, Julia Faix, Jan Ngezahayo, Anaclet Bleich, André Heisterkamp, Alexander Buettner, Manuela Kalies, Stefan Epithelial restitution in 3D - Revealing biomechanical and physiochemical dynamics in intestinal organoids via fs laser nanosurgery |
title | Epithelial restitution in 3D - Revealing biomechanical and physiochemical dynamics in intestinal organoids via fs laser nanosurgery |
title_full | Epithelial restitution in 3D - Revealing biomechanical and physiochemical dynamics in intestinal organoids via fs laser nanosurgery |
title_fullStr | Epithelial restitution in 3D - Revealing biomechanical and physiochemical dynamics in intestinal organoids via fs laser nanosurgery |
title_full_unstemmed | Epithelial restitution in 3D - Revealing biomechanical and physiochemical dynamics in intestinal organoids via fs laser nanosurgery |
title_short | Epithelial restitution in 3D - Revealing biomechanical and physiochemical dynamics in intestinal organoids via fs laser nanosurgery |
title_sort | epithelial restitution in 3d - revealing biomechanical and physiochemical dynamics in intestinal organoids via fs laser nanosurgery |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10585398/ https://www.ncbi.nlm.nih.gov/pubmed/37867948 http://dx.doi.org/10.1016/j.isci.2023.108139 |
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