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Sequential bottom-up assembly of mechanically stabilized synthetic cells by microfluidics

MPS-Authors
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Weiss,  Marian
Cellular Biophysics, Max Planck Institute for Medical Research, Max Planck Society;

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Frohnmayer,  Johannes Patrick
Cellular Biophysics, Max Planck Institute for Medical Research, Max Planck Society;
Biophysical Chemistry, Institute of Physical Chemistry, University of Heidelberg, 69120 Heidelberg, Germany;

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Benk,  Lucia Theresa
Cellular Biophysics, Max Planck Institute for Medical Research, Max Planck Society;

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Haller,  Barbara
Cellular Biophysics, Max Planck Institute for Medical Research, Max Planck Society;

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Janiesch,  Jan-Willi
Cellular Biophysics, Max Planck Institute for Medical Research, Max Planck Society;
Biophysical Chemistry, Institute of Physical Chemistry, University of Heidelberg, 69120 Heidelberg, Germany;

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Platzman,  Ilia
Cellular Biophysics, Max Planck Institute for Medical Research, Max Planck Society;
Biophysical Chemistry, Institute of Physical Chemistry, University of Heidelberg, 69120 Heidelberg, Germany;

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Spatz,  Joachim P.
Cellular Biophysics, Max Planck Institute for Medical Research, Max Planck Society;
Biophysical Chemistry, Institute of Physical Chemistry, University of Heidelberg, 69120 Heidelberg, Germany;

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Citation

Weiss, M., Frohnmayer, J. P., Benk, L. T., Haller, B., Janiesch, J.-W., Heitkamp, T., et al. (2018). Sequential bottom-up assembly of mechanically stabilized synthetic cells by microfluidics. Nature Materials, 17, 89-96. doi:10.1038/nmat5005.


Cite as: http://hdl.handle.net/11858/00-001M-0000-002E-0AA2-B
Abstract
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.