Abstract
(1) Background: giant vesicles (GVs) are widely employed as models for studying physicochemical properties of bio-membranes and artificial cell construction due to their similarities to natural cell membranes. Considering the critical roles of GVs, various methods have been developed to prepare them. Notably, the water-in-oil (w/o) inverted emulsion-transfer method is reported to be the most promising, owning to the relatively higher productivity and better encapsulation efficiency of biomolecules. Previously, we successfully established an improved approach to acquire detailed information of 1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC)-derived GVs with imaging flow cytometry (IFC); (2) Methods: we prepared GVs with different lipid compositions, including phosphatidylcholines (PCs), phosphatidylethanolamines (PEs), and PC/PE mixtures by w/o inverted emulsion methods. We comprehensively compared the yield, purity, size, and encapsulation efficiency of the resulting vesicles; (3) Results: the relatively higher productivities of GVs could be obtained from POPC, 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dilauroyl-sn-glycero-3-phosphoethanolamine (DLPE), DOPC: DLPE (7:3), and POPC: DLPE (6:4) pools. Furthermore, we also demonstrate that these GVs are stable during long term preservation in 4 °C. (4) Conclusions: our results will be useful for the analytical study of GVs and GV-based applications.
Highlights
To understand the organization and dynamics of lipid bilayers, research on giant vesicles (GVs) as an artificial cellular membrane model has gained considerable interest in recent decades [1,2,3,4,5,6]
Continuing that effort in this study, we report the preparation of GVs with different lipid compositions such as PC, PE, and PC-PE mixtures in different ratios by the water-inoil inverted emulsion method
Using imaging flow cytometry (IFC) to Analyze GVs Produced by the w/o Emulsion Transfer Method
Summary
To understand the organization and dynamics of lipid bilayers, research on giant vesicles (GVs) as an artificial cellular membrane model has gained considerable interest in recent decades [1,2,3,4,5,6]. Compared to the methods mentioned above, phase transfer methods such as the water-in-oil (w/o) emulsion transfer, has shown significant advantages, since it does not need specialized equipment, is of straightforward implementation, acceptable encapsulation efficiency, and high yield of uni-lamellar vesicles [15,16], one drawback is that there is a possible presence of oil trapped within the bilayers depending on the protocols and reagent used, which might influence membrane mechanical properties [17,25,26] This method can produce GVs with a wide range of lipid compositions, e.g., phosphatidylcholines (PCs) and phosphatidylethanolamines (PEs), which are usually employed for constructing the artificial cell membrane [6,27,28]. GVs comprehensive from w/o emulsion method show high stability overwe long incubation of GVs significantly high encapsulation efficiency
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