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...

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Bibliographic Details
Published in:	iScience Vol. 26; no. 11; p. 108139
Main Authors:	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
Format:	Journal Article
Language:	English
Published:	Elsevier Inc 17-11-2023 Elsevier
Subjects:	Bioengineering Cell biology Molecular physiology Optical imaging Bioengineering Cell biology Optical imaging Molecular physiology
Online Access:	Get full text
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Summary:	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. [Display omitted] •Lesions in crypt bases of colonoids are closed by the flattening of adjacent cells•Epithelial cell layer changes viscoelastic properties during epithelial restitution•Contractile forces of the cytoskeleton are indispensable for wound healing•Single cell ablation in crypt base leads to altered Calcium- and YAP1-signaling Optical imaging; Molecular physiology; Bioengineering; Cell biology
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Lead contact
ISSN:	2589-0042 2589-0042
DOI:	10.1016/j.isci.2023.108139