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Free keywords:
synthetic cell, droplet-based microfluidics, water-in-oil-droplets, giant unilamellar vesicles, fluorosurfactants, compartments
Abstract:
ABSTRACT: In this manuscript, we introduce a simple, off-the-shelf approach for the on-demand creation of giant unilamellar vesicles (GUVs) or multicompartment synthetic cell model systems in a high-throughput manner. To achieve this, we use microfluidics to encapsulate small unilamellar vesicles in block-copolymer surfactant-stabilized water-in-oil droplets. By tuning the charge of the inner droplet interface, adsorption of lipids can either be inhibited, leading to multicompartment systems, or induced, leading to the formation of a droplet-stabilized GUV. To control the charge density, we formed droplets using different molar ratios of an uncharged PEG-based fluorosurfactant and a negatively-charged PFPE carboxylic acid fluorosurfactant (Krytox). We systematically studied the transition from a multicompartment system to 3D-supported lipid bilayers as a function of lipid charge and Krytox concentration using confocal fluorescence microscopy and interfacial tension measurements. Moreover, we showed and characterized the release of assembled GUVs from the surfactant shell and the oil phase into a physiological buffer, an approach for the formation of GUVs with improved fea-tures that is high in yield compared to standard GUV formation methods. This widely applica-ble microfluidic-based technology will increase the scope of usage for GUVs as adaptable cell-like compartments in bottom-up synthetic biology applications and beyond.
