Evaluation of dextran(ethylene glycol) hydrogel films for giant unilamellar lipid vesicle production and their application for the encapsulation of polymersomes

Abstract

Giant Unilamellar Vesicles (GUVs) prepared from phospholipids are becoming popular membrane model systems for use in biophysical studies. The quality, size and yield of GUVs depend on the preparation method used to obtain them. In this study, hydrogels consisting of dextran polymers crosslinked by poly(ethylene glycol) (DexPEG) were used as hydrophilic frameworks for the preparation of vesicle suspensions under physiological ionic strength conditions. A comparative study was conducted using hydrogels with varied physicochemical properties to evaluate their performance for GUV production. The prepared GUVs were quantified by flow cytometry using the Coulter Principle to determine yield and size distribution. We find that hydrogels of lower mechanical strength, increased swellability and decreased lipid interaction favour GUV production, while their resulting size is determined by the surface roughness of the hydrogel film. Moreover, we embedded polymersomes into the crosslinked hydrogel network, creating a DexPEG – polymersomes hybrid film. The re-hydration of lipids on those hybrid substrates led to production of GUVs and the efficient encapsulation of polymersomes in the lumen of GUVs.

Controlled Peptide-Mediated Vesicle Fusion Assessed by Simultaneous Dual-Colour Time-Lapsed Fluorescence Microscopy

“…The corresponding intensities of the donors are shown in panel C. Representative images of GUVs are shown on panel D. The emission of the donors was recorded at 515 nm. (2020) 10:3087 | https://doi.org/10.1038/s41598-020-59926-z www.nature.com/scientificreports www.nature.com/scientificreports/ DexPEG substrates allows for the growth of GUVs under physiological conditions in good yields 59 . The lipid film was deposited on DexPEG substrates and hydrated at room temperature with phosphate buffer solution (PBS, pH = 7.4), containing CaCl 2 (1 mM), MgCl 2 (0.5 mM) and sucrose (200 mM).…”

The membrane transporter lactose permease increases lipid bilayer bending rigidity

“…LacY GUVs were prepared from LacY LUVs in a similar way to the partial dehydration and electroformation method ( 16), but the electroswelling step was replaced by the use of DexPEG hydrogels for GUV generation, which has been proven to be compatible with the use of anionic lipids and physiological ionic strength conditions (35,46) required to preserve biological protein activity in the lipid membrane. For instance, lack of functionality in bacteriorhodopsin was found when the reconstitution was performed with the partial dehydration and electroswelling method just in water (16).…”

Furfuryl‐ and Maleimido Polysaccharides: Synthetic Strategies Toward Functional Biomaterials

“…Products with DS values of 0.10 to 0.26 were obtained as calculated from the 1 H NMR signals of maleimide and glucosidic protons . Moreover, N , N ′‐diisopropylcarbodiimide (DIC) was used to perform the esterification of dextran with N ‐maleoyl β‐alanine in DMSO . Catalytic amounts of 4‐(dimethylamino) pyridinium 4‐toluenesulfonate (DPTS) were added to the reaction mixture ( Table 3 , Figure 6 ).…”

“…The drop‐casting on thiolated glass slides yielded hydrogel films, which were coated with phospholipid solutions to produce giant unilamellar vesicles (GUVs) upon film rehydration. The study is interesting for membrane model systems in biophysics allowing the encapsulation of biomolecular cargos and particles . The authors believe that GUVs can be used to design cell models providing insights into cellular structure in context with the origin of life.…”