Abstract
All living organisms sense mechanical forces. Engineering mechanosensitive artificial cell through bottom-up in vitro reconstitution offers a way to understand how mixtures of macromolecules assemble and organize into a complex system that responds to forces. We use stable double emulsion droplets (aqueous/oil/aqueous) to prototype mechanosensitive artificial cells. In order to demonstrate mechanosensation in artificial cells, we develop a novel microfluidic device that is capable of trapping double emulsions into designated chambers, followed by compression and aspiration in a parallel manner. The microfluidic device is fabricated using multilayer soft lithography technology, and consists of a control layer and a deformable flow channel. Deflections of the PDMS membrane above the main microfluidic flow channels and trapping chamber array are independently regulated pneumatically by two sets of integrated microfluidic valves. We successfully compress and aspirate the double emulsions, which result in transient increase and permanent decrease in oil thickness, respectively. Finally, we demonstrate the influx of calcium ions as a response of our mechanically activated artificial cell through thinning of oil. The development of a microfluidic device to mechanically activate artificial cells creates new opportunities in force-activated synthetic biology.
Highlights
In order to demonstrate mechanosensation in artificial cells, we develop a novel microfluidic device that is capable of trapping double emulsions into designated chambers, followed by compression and aspiration in a parallel manner
Microfluidics technology has been widely adopted in single cell mechanobiology studies where the effect of different modes of mechanical forces on cellular functions have been investigated[29,30,31,32,33,34]
To identify the ideal spinning speed and membrane thickness for the microfluidic device, we considered two criteria, which were the strength of PDMS bonding and the strength of the membrane from rupturing
Summary
In order to demonstrate mechanosensation in artificial cells, we develop a novel microfluidic device that is capable of trapping double emulsions into designated chambers, followed by compression and aspiration in a parallel manner. Bottom-up reconstitution offers a way to peel away cellular complexity through building from cellular components to understand how macromolecules assemble and organize into complex structures or generate emergent behaviors[3,4], and building bio-inspired artificial cells where inherent complexities of living cells are stripped away holds tremendous promise in a broad range of applications[5,6]. Building artificial cell systems directly with lipid bilayer has challenged many research groups, mainly due to low stability of artificial cells This has motivated us to use double emulsion droplets (aqueous/oil/aqueous) as an alternative model system to prototype artificial cells. The capability and controllability of microfluidics offer a versatile platform for engineering and studying mechanically activated artificial cells
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