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Sci Rep. 2020 Mar 16;10(1):4824. doi: 10.1038/s41598-020-61655-2.

Facile generation of giant unilamellar vesicles using polyacrylamide gels.

Author information

1
University Hospital Balgrist, University of Zurich, Zürich, Switzerland.
2
Institute for Biomechanics, ETH Zurich, Zürich, Switzerland.
3
Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, USA.
4
MRC Laboratory of Molecular Biology, Cambridge, United Kingdom.
5
Laboratory of Physical Chemistry, ETH Zurich, Zürich, Switzerland.
6
Center for Microscopy and Image Analysis, University of Zurich, Zurich, Switzerland.
7
Flow Cytometry Facility, University of Zurich, Zurich, Switzerland.
8
NCCR Chemical Biology, Department of Biochemistry, University of Geneva, Geneva, Switzerland.
9
University Hospital Balgrist, University of Zurich, Zürich, Switzerland. unai.silvan@hest.ethz.ch.
10
Institute for Biomechanics, ETH Zurich, Zürich, Switzerland. unai.silvan@hest.ethz.ch.

Abstract

Giant unilamellar vesicles (GUVs) are model cell-sized systems that have broad applications including drug delivery, analysis of membrane biophysics, and synthetic reconstitution of cellular machineries. Although numerous methods for the generation of free-floating GUVs have been established over the past few decades, only a fraction have successfully produced uniform vesicle populations both from charged lipids and in buffers of physiological ionic strength. In the method described here, we generate large numbers of free-floating GUVs through the rehydration of lipid films deposited on soft polyacrylamide (PAA) gels. We show that this technique produces high GUV concentrations for a range of lipid types, including charged ones, independently of the ionic strength of the buffer used. We demonstrate that the gentle hydration of PAA gels results in predominantly unilamellar vesicles, which is in contrast to comparable methods analyzed in this work. Unilamellarity is a defining feature of GUVs and the generation of uniform populations is key for many downstream applications. The PAA method is widely applicable and can be easily implemented with commonly utilized laboratory reagents, making it an appealing platform for the study of membrane biophysics.

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