Abstract

Giant unilamellar phospholipid vesicles are attractive starting points for constructing minimal living cells from the bottom-up. Their membranes are compatible with many physiologically functional modules and act as selective barriers, while retaining a high morphological flexibility. However, their spherical shape renders them rather inappropriate to study phenomena that are based on distinct cell shape and polarity, such as cell division. Here, a microscale device based on 3D printed protein hydrogel is introduced to induce pH-stimulated reversible shape changes in trapped vesicles without compromising their free-standing membranes. Deformations of spheres to at least twice their aspect ratio, but also toward unusual quadratic or triangular shapes can be accomplished. Mechanical force induced by the cages to phase-separated membrane vesicles can lead to spontaneous shape deformations, from the recurrent formation of dumbbells with curved necks between domains to full budding of membrane domains as separate vesicles. Moreover, shape-tunable vesicles are particularly desirable when reconstituting geometry-sensitive protein networks, such as reaction-diffusion systems. In particular, vesicle shape changes allow to switch between different modes of self-organized protein oscillations within, and thus, to influence reaction networks directly by external mechanical cues.

Highlights

  • Giant unilamellar phospholipid vesicles are attractive starting points for on the other hand.[3]

  • But still being compatible with these established protocols, our 3D BSA protein hydrogel GUVs traps were fabricated in a layerby-layer procedure via two-photon polymerization process, using Rose bengal as the photoinitiator for BSA monomers (Scheme S1, Supporting Information)

  • We have developed a new toolbox for mechanical manipulation of GUVs—model membrane vesicles that constitute the basis for the engineering of advanced protocells and that should ideally be subject to defined shape transformations

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Summary

Chip design

Tissue scaffolds[17] and generating smart 4D stimuli-responsive microactuators.[16,18]. We varied and expanded this technology toward the goal of selectively trapping GUVs within a customized 3D printed BSA hydrogel chip, and dynamically inducing structural anisotropy by applying external pH stimuli to the gel. 3D printed protein hydrogel can be designed as microchambers in appropriate sizes for capturing GUVs. The variable protein hydrogel structure acts as a geometrical cue to establish synthetic cell polarity in vitro by compressing vesicles into different shapes upon pH stimuli. The variable protein hydrogel structure acts as a geometrical cue to establish synthetic cell polarity in vitro by compressing vesicles into different shapes upon pH stimuli This spatially well-defined microenvironment can mimic the dynamic native cell matrix, allowing us to investigate how synthetic cells react to and interact with external mechanical cues

Results and Discussion
Conclusion
Conflict of Interest
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