2-photon laser printing to mechanically stimulate multicellular systems in 3D
Most biological activities take place in 3D environments, where cells communicate with each other in various directions and are located in a defined, often microstructured, space. To investigate the effect of defined cyclic mechanical forces on a multicellular system, we develop a sub-millimeter sized stretching device for mechanical stimulation of a structurally restricted, soft multicellular microenvironment. For the stretching device, a multimaterial 3D microstructure made of PDMS and gelatine-based hydrogel is printed via 2-photon polymerization (2PP) method. The printed structures are first characterized microscopically and mechanically to study the effect of different printing parameters. With 2PP, organotypic cell cultures are then directly printed into the hydrogel structures to achieve true 3D cell culture systems. These are mechanically stimulated with a cantilever by indenting the stretching device at a defined point. As a most important result, the cells in the 3D organotypic cell culture change morphology and actin orientation when exposed to cyclic mechanical stretch, even within short timescales of just 30 minutes. As a proof of concept, we encapsulated a Medaka retinal organoid in the same structure to demonstrate that even preformed organoids can be stimulated by our method. The results demonstrate the power of 2PP to manufacturing multifunctional soft devices for mechanically controlling multicellular systems at micrometer resolution and thus mimicking mechanical stress situations, as they occurin vivo..
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Preprint| Erscheinungsjahr: |
2023 |
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| Erschienen: |
2023 |
| Enthalten in: |
bioRxiv.org - (2023) vom: 29. Dez. Zur Gesamtaufnahme - year:2023 |
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| Sprache: |
Englisch |
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| Beteiligte Personen: |
Colombo, Federico [VerfasserIn] |
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| Links: |
Volltext [kostenfrei] |
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| doi: |
10.1101/2023.12.23.573049 |
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XBI041980697 |
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| 520 | |a Most biological activities take place in 3D environments, where cells communicate with each other in various directions and are located in a defined, often microstructured, space. To investigate the effect of defined cyclic mechanical forces on a multicellular system, we develop a sub-millimeter sized stretching device for mechanical stimulation of a structurally restricted, soft multicellular microenvironment. For the stretching device, a multimaterial 3D microstructure made of PDMS and gelatine-based hydrogel is printed via 2-photon polymerization (2PP) method. The printed structures are first characterized microscopically and mechanically to study the effect of different printing parameters. With 2PP, organotypic cell cultures are then directly printed into the hydrogel structures to achieve true 3D cell culture systems. These are mechanically stimulated with a cantilever by indenting the stretching device at a defined point. As a most important result, the cells in the 3D organotypic cell culture change morphology and actin orientation when exposed to cyclic mechanical stretch, even within short timescales of just 30 minutes. As a proof of concept, we encapsulated a Medaka retinal organoid in the same structure to demonstrate that even preformed organoids can be stimulated by our method. The results demonstrate the power of 2PP to manufacturing multifunctional soft devices for mechanically controlling multicellular systems at micrometer resolution and thus mimicking mechanical stress situations, as they occurin vivo. | ||
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