Xu Boying, Ding Jinquan, Xu Jian, Yomo Tetsuya
Laboratory of Biology and Information Science, Biomedical Synthetic Biology Research Center, School of Life Sciences, East China Normal University, Shanghai 200062, China.
School of Software Engineering, East China Normal University, Shanghai 200062, China.
Life (Basel). 2021 Mar 10;11(3):223. doi: 10.3390/life11030223.
(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.
(1) 背景：由于巨型囊泡（GVs）与天然细胞膜相似，它们被广泛用作研究生物膜物理化学性质和人工细胞构建的模型。考虑到GVs的关键作用，已经开发了各种方法来制备它们。值得注意的是，油包水（w/o）反相乳液转移法据报道是最有前景的，因为其生物分子的生产率相对较高且封装效率更好。此前，我们成功建立了一种改进方法，通过成像流式细胞术（IFC）获取1-棕榈酰-2-油酰-sn-甘油-3-磷酸胆碱（POPC）衍生的GVs的详细信息；(2) 方法：我们通过w/o反相乳液法制备了具有不同脂质组成的GVs，包括磷脂酰胆碱（PCs）、磷脂酰乙醇胺（PEs）以及PC/PE混合物。我们全面比较了所得囊泡的产量、纯度、大小和封装效率；(3) 结果：从POPC、1,2-二油酰-sn-甘油-3-磷酸胆碱（DOPC）、1,2-二月桂酰-sn-甘油-3-磷酸乙醇胺（DLPE）、DOPC:DLPE（7:3）和POPC:DLPE（6:4）组合中可以获得相对较高生产率的GVs。此外，我们还证明这些GVs在4°C长期保存期间是稳定的。(4) 结论：我们的结果将有助于GVs的分析研究和基于GVs的应用。
