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Cell-Cell Separation Device: measurement of intercellular detachment forces

  • Whether at the intramolecular or cellular scale in organisms, cell-cell adhesion adapt to external mechanical cues arising from the static environment of cells and from dynamic interactions between neighboring cells. Cell-cell adhesions need to resist detachment forces to secure the integrity and internal organization of organisms. In the past, various techniques have been developed to characterize adhesion properties of molecules and cells in vitro, and to understand how cells sense and probe their environment. Atomic force microscopy and dual-pipette aspiration, where cells are mainly present in suspension, are common methods for studying detachment forces of cell-cell adhesions. How cell-cell adhesion forces are developed for adherent and environment-adapted cells, however, is less clear. Here, we designed the Cell-Cell Separation Device (CC-SD), a microstructured substrate that measures both intercellular forces and external stresses of cells towards the matrix. The design is based on micropillar arrays originally designed for cell traction-force measurements. We designed PDMS micropillar-blocks, to which cells could adhere and be able to connect to each other across the gap. Controlled stretching of the whole substrate changed the distance between blocks and increased gap size. That allowed us to apply strains to cell-cell contacts, eventually leading to cell-cell adhesion detachment, which was measured by pillar deflections. The CC-SD provided an increase of the gap between the blocks of up to 2.4-fold, which was sufficient to separate substrate-attached cells with fully developed F-actin network. Simultaneously measured pillar deflections allowed us to address cellular response to the intercellular strain applied. The CC-SD thus opens up possibilities for the analysis of intercellular force detachments and sheds light on the robustness of cell-cell adhesions in dynamic processes in tissue development.

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Metadaten
Author:Julia Eckert, Volha Matylitskaya, Stephan Kasemann, Stefan Partel, Thomas Schmidt
DOI:https://doi.org/10.1101/2023.03.16.532950
Document Type:Preprint
Language:English
Year of publication:2023
Release Date:2024/01/31
Number of pages:18
Note:
bioRxiv - The Preprint Server for Biology
Organisationseinheit:Forschung / Forschungszentrum Mikrotechnik
DDC classes:500 Naturwissenschaften und Mathematik
Peer review:wiss. Beitrag, nicht peer-reviewed
Publicationlist:Kasemann, Stephan
Matylitskaya, Volha
Partel, Stefan