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Artificial hybrid cells created by ns-pulse mediated electrofusion

STSM by Christoph Huber, Early-Stage Researcher

Period of mission: from 14/04/2014 to 18/04/2014

Home institution: Institute of Synthetic Bioarchitectures, Department of Nanobiotechnology, University of Natural Resources and Life Sciences, Vienna, Austria

Host institution: Laboratory of Biocybernetics, Faculty of Electrical Engineering, University of Ljubljana, Ljubljana, Slovenia

Classical electrofusion is considered to be an established technique to fuse cells of similar sizes. It was shown recently that fusion of cells of different sizes is also possible but the application of ns pulses instead of ms- or µs pulses is necessary. This new possibility gave rise to the idea of fusing artificial vesicles, built of amphiphilic block copolymers, with living cells. The so called polymersomes offer a broad variety of physical and chemical properties, determined by the type of polymers used as their building blocks. Polymersomes can work as a shuttle system for molecules filled in their inner compartment, as their permeability is typically lower than the permeability of liposomes and can be precisely tuned by the chemical structure and/or the molecular weight of the used polymers. Moreover polymersomes can work as a support for transmembrane proteins, as these proteins can only maintain their functionality if embedded in the appropriate environment. During the STSM incipient experiments were made to investigate the ability to electroporate polymersomes in principal. It turned out that ns pulses were not strong enough to lead to sufficient pore formation to be investigated in a classical fluorescence microscope. This could be overcome by the application of µs pulses and was monitored by the leakage of fluorophores trapped in the inner compartment of the polymersomes. Moreover it could be demonstrated that close contact between cells and polymersomes can be established by dielectrophoresis, which is an essential prerequisite for successful fusion. To demonstrate fusion between cells and polymersomes further experiments are already planned.


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