Sequential bottom-up assembly of mechanically stabilized synthetic cells by microfluidics
Authors: Weiss M, Frohnmayer JP, Benk LT, Haller B, Janiesch JW, Heitkamp T, Börsch M, Lira RB, Dimova R, Lipowsky R, Bodenschatz E, Baret JC, Vidakovic-Koch T, Sundmacher K, Platzman I, Spatz JP
CellNetworks People: Spatz Joachim
Journal: Nat Mater. 2018 Jan;17(1):89-96. doi: 10.1038/nmat5005.

Compartments for the spatially and temporally controlled assembly of biological processes are essential towards cellular life. Synthetic mimics of cellular compartments based on lipid-based protocells lack the mechanical and chemical stability to allow their manipulation into a complex and fully functional synthetic cell. Here, we present a high-throughput microfluidic method to generate stable, defined sized liposomes termed 'droplet-stabilized giant unilamellar vesicles (dsGUVs)'. The enhanced stability of dsGUVs enables the sequential loading of these compartments with biomolecules, namely purified transmembrane and cytoskeleton proteins by microfluidic pico-injection technology. This constitutes an experimental demonstration of a successful bottom-up assembly of a compartment with contents that would not self-assemble to full functionality when simply mixed together. Following assembly, the stabilizing oil phase and droplet shells are removed to release functional self-supporting protocells to an aqueous phase, enabling them to interact with physiologically relevant matrices.