Electroformation of giant liposomes in microfluidic channels

Kuribayashi, Kaori; Tresset, Guillaume; Coquet, P.; Fujita, Hiroyuki; Takeuchi, Shoji

doi:10.1088/0957-0233/17/12/s01

Cited by 66 publications

(60 citation statements)

References 21 publications

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“…To date, standard methods have been used for preparation of giant vesicles [13], [14], that encapsulate substances; however, few reports have described the encapsulation of micrometer-sized substances into giant vesicles at high volume fractions [15]. Recently, the water-in-oil (w/o) emulsion centrifugation method has been developed.…”

Section: Introductionmentioning

confidence: 99%

Introducing Micrometer-Sized Artificial Objects into Live Cells: A Method for Cell–Giant Unilamellar Vesicle Electrofusion

et al. 2014

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Here, we report a method for introducing large objects of up to a micrometer in diameter into cultured mammalian cells by electrofusion of giant unilamellar vesicles. We prepared GUVs containing various artificial objects using a water-in-oil (w/o) emulsion centrifugation method. GUVs and dispersed HeLa cells were exposed to an alternating current (AC) field to induce a linear cell–GUV alignment, and then a direct current (DC) pulse was applied to facilitate transient electrofusion. With uniformly sized fluorescent beads as size indexes, we successfully and efficiently introduced beads of 1 µm in diameter into living cells along with a plasmid mammalian expression vector. Our electrofusion did not affect cell viability. After the electrofusion, cells proliferated normally until confluence was reached, and the introduced fluorescent beads were inherited during cell division. Analysis by both confocal microscopy and flow cytometry supported these findings. As an alternative approach, we also introduced a designed nanostructure (DNA origami) into live cells. The results we report here represent a milestone for designing artificial symbiosis of functionally active objects (such as micro-machines) in living cells. Moreover, our technique can be used for drug delivery, tissue engineering, and cell manipulation.

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Section: Introductionmentioning

confidence: 99%

Introducing Micrometer-Sized Artificial Objects into Live Cells: A Method for Cell–Giant Unilamellar Vesicle Electrofusion

et al. 2014

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“…Both micron-and nanometer-sized liposomes have been produced in microfluidic devices, however these methods suffer from either polydisperse size populations or poor encapsulation efficiency. 14,15 Other methods have been presented which implement an ink-jet powered technique that produces monodisperse lipid vesicles, but the vesicles generated are hundreds of micrometers in diameter. 16 Another method uses a lipid bubble blowing process but is limited in its ability to vary the size of the vesicles generated.…”

Section: Introductionmentioning

confidence: 99%

Stable, biocompatible lipid vesicle generation by solvent extraction-based droplet microfluidics

et al. 2011

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In this paper, we present a microfluidic platform for the continuous generation of stable, monodisperse lipid vesicles 20-110 lm in diameter. Our approach utilizes a microfluidic flow-focusing droplet generation design to control the vesicle size by altering the system's fluid flow rates to generate vesicles with narrow size distribution. Double emulsions are first produced in consecutive flow-focusing channel geometries and lipid membranes are then formed through a controlled solvent extraction process. Since no strong solvents are used in the process, our method allows for the safe encapsulation and manipulation of an assortment of biological entities, including cells, proteins, and nucleic acids. The vesicles generated by this method are stable and have a shelf life of at least 3 months. Here, we demonstrate the cell-free in vitro synthesis of proteins within lipid vesicles as an initial step towards the development of an artificial cell.

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“…By using of the analysing device combination with quantitative unilamellar giant liposome formation device in the microfluidics [6] will provide a sophisticated tools for artificial cell model research.…”

Section: Resultsmentioning

confidence: 99%

Giant liposome sorting/collection device: For individual analysis of artificial cell-models

Nomura

Liu

Chen

et al. 2009

2009 International Symposium on Micro-NanoMechatronics and Human Science

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In this report, we will present a novel liposome handling method on microfluidic channel. For the quantitative analysis of individual giant liposomes, a microfluidic device with multi-pillar was designed. Liposomes suspension from neutral phospholipids prepared by natural swelling method was introduced into the separation device. As a result, giant liposomes are safely trapped in the microscope viewfield along with the internal pillar within a weak flow. In the condition, small solutes (excess dye molecules, small liposomes) are washed away through the pillars. This system should be useful for time-course analysis of individual cell-sized liposomes.

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Electroformation of giant liposomes in microfluidic channels

Cited by 66 publications

References 21 publications

Introducing Micrometer-Sized Artificial Objects into Live Cells: A Method for Cell–Giant Unilamellar Vesicle Electrofusion

Introducing Micrometer-Sized Artificial Objects into Live Cells: A Method for Cell–Giant Unilamellar Vesicle Electrofusion

Stable, biocompatible lipid vesicle generation by solvent extraction-based droplet microfluidics

Giant liposome sorting/collection device: For individual analysis of artificial cell-models

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