Microfluidic Handling and Analysis of Giant Vesicles for Use as Artificial Cells: A Review

Robinson, Tom

doi:10.1002/adbi.201800318

Cited by 46 publications

(44 citation statements)

References 107 publications

(219 reference statements)

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“…[126] Recently, microsized vesicles (mainly giant unilamellar vesicles) have surged to a new role as ideal models of artificial cells and cell membrane. [132] The advantages derived from the use of microfluidics technique for their production reside mainly in the degree of control achievable on size, lamellarity, membrane composition, and payload. [131,133,134] At the same time it is possible to produce high number of such vesicles, reducing the overall costs.…”

Section: Microvesicles: From Polymerosomes To Artificial Cellsmentioning

confidence: 99%

“…[137] Interestingly, by mechanical compressing the giant unilamellar vesicles, Robinson et al observed a spontaneous rearrangement of the lipid domains culminating into a the formation of bigger domains. [132] Artificial cells and artificial cell membranes help in the breaking down of the complex cellular machine, one mechanism at the time to better understand their function. [133,138] However, the complexity of the cellular environment has proven difficult to recreate either in terms of membrane features (as discussed above) or in terms of intracellular organelles complexity.…”

Section: Microvesicles: From Polymerosomes To Artificial Cellsmentioning

confidence: 99%

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Microfluidics for Production of Particles: Mechanism, Methodology, and Applications

Liu

Fontana

Python

et al. 2019

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In the past two decades, microfluidics‐based particle production is widely applied for multiple biological usages. Compared to conventional bulk methods, microfluidic‐assisted particle production shows significant advantages, such as narrower particle size distribution, higher reproducibility, improved encapsulation efficiency, and enhanced scaling‐up potency. Herein, an overview of the recent progress of the microfluidics technology for nano‐, microparticles or droplet fabrication, and their biological applications is provided. For both nano‐, microparticles/droplets, the previously established mechanisms behind particle production via microfluidics and some typical examples during the past five years are discussed. The emerging interdisciplinary technologies based on microfluidics that have produced microparticles or droplets for cellular analysis and artificial cells fabrication are summarized. The potential drawbacks and future perspectives are also briefly discussed.

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Section: Microvesicles: From Polymerosomes To Artificial Cellsmentioning

confidence: 99%

Section: Microvesicles: From Polymerosomes To Artificial Cellsmentioning

confidence: 99%

Microfluidics for Production of Particles: Mechanism, Methodology, and Applications

Liu

Fontana

Python

et al. 2019

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“…GUVs are considered the gold standard of cellular mimics owing to their similarity in size to that of a mammalian cell. Moreover, with advances in technologies, such as microfluidics for their production and handling, GUVs are looking like an increasingly more attractive option.…”

Section: Introductionmentioning

confidence: 99%

Optimization of the Inverted Emulsion Method for High‐Yield Production of Biomimetic Giant Unilamellar Vesicles

et al. 2019

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In the field of bottom‐up synthetic biology, lipid vesicles provide an important role in the construction of artificial cells. Giant unilamellar vesicles (GUVs), due to their membrane's similarity to natural biomembranes, have been widely used as cellular mimics. So far, several methods exist for the production of GUVs with the possibility to encapsulate biological macromolecules. The inverted emulsion‐based method is one such technique, which has great potential for rapid production of GUVs with high encapsulation efficiencies for large biomolecules. However, the lack of understanding of various parameters that affect production yields has resulted in sparse adaptation within the membrane and bottom‐up synthetic biology research communities. Here, we optimize various parameters of the inverted emulsion‐based method to maximize the production of GUVs. We demonstrate that the density difference between the emulsion droplets, oil phase, and the outer aqueous phase plays a crucial role in vesicle formation. We also investigated the impact that centrifugation speed/time, lipid concentration, pH, temperature, and emulsion droplet volume has on vesicle yield and size. Compared to conventional electroformation, our preparation method was not found to significantly alter the membrane mechanical properties. Finally, we optimize the parameters to minimize the time from workbench to microscope and in this way open up the possibility of time‐sensitive experiments. In conclusion, our findings will promote the usage of the inverted emulsion method for basic membrane biophysics studies as well as the development of GUVs for use as future artificial cells.

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“…For this reason, it will be interesting to observe how multiple vesicle systems respond to external factors: not only compression, as shown here on single artificial cells, but with osmotic gradients, changes in heat, and also applied shear stresses. We are therefore developing systems to model proto‐tissues by using high‐capacity microfluidic vesicle capture with this future goal in mind …”

Section: Resultsmentioning

confidence: 99%

Observations of Membrane Domain Reorganization in Mechanically Compressed Artificial Cells

Robinson

Dittrich

2019

ChemBioChem

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Giant unilamellar vesicles (GUVs) are considered to be the gold standard for assembling artificial cells from the bottom up. In this study, we investigated the behavior of such biomimetic vesicles as they were subjected to mechanical compression. A microfluidic device is presented that comprises a trap to capture GUVs and a microstamp that is deflected downwards to mechanically compress the trapped vesicle. After characterization of the device, we show that single‐phase GUVs can be controllably compressed to a high degree of deformation (D=0.40) depending on the pressure applied to the microstamp. A permeation assay was implemented to show that vesicle bursting is prevented by water efflux. Next, we mechanically compressed GUVs with co‐existing liquid‐ordered and liquid‐disordered membrane phases. Upon compression, we observed that the normally stable lipid domains reorganized themselves across the surface and fused into larger domains. This phenomenon, observed here in a model membrane system, not only gives us insights into how the multicomponent membranes of artificial cells behave, but might also have interesting consequences for the role of lipid rafts in biological cells that are subjected to compressive forces in a natural environment.

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Microfluidic Handling and Analysis of Giant Vesicles for Use as Artificial Cells: A Review

Cited by 46 publications

References 107 publications

Microfluidics for Production of Particles: Mechanism, Methodology, and Applications

Microfluidics for Production of Particles: Mechanism, Methodology, and Applications

Optimization of the Inverted Emulsion Method for High‐Yield Production of Biomimetic Giant Unilamellar Vesicles

Observations of Membrane Domain Reorganization in Mechanically Compressed Artificial Cells

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