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氟化纳米颗粒辅助形成巨型单层囊泡。

Formation of Giant Unilamellar Vesicles Assisted by Fluorinated Nanoparticles.

机构信息

Mechatronics, Biostatistics and Sensors (MeBioS) at KU Leuven, Willem de Croylaan 42, 3001, Leuven, Belgium.

Soft Matter, Rheology and Technology (SMaRT) at KU Leuven, Celestijnenlaan 200J, 3000, Leuven, Belgium.

出版信息

Adv Sci (Weinh). 2023 Dec;10(34):e2302461. doi: 10.1002/advs.202302461. Epub 2023 Oct 9.

Abstract

In the quest to produce artificial cells, one key challenge that remains to be solved is the recreation of a complex cellular membrane. Among the existing models, giant unilamellar vesicles (GUVs) are particularly interesting due to their intrinsic compartmentalisation ability and their resemblance in size and shape to eukaryotic cells. Many techniques have been developed to produce GUVs all having inherent advantages and disadvantages. Here, the authors show that fluorinated silica nanoparticles (FNPs) used to form Pickering emulsions in a fluorinated oil can destabilise lipid nanosystems to template the formation of GUVs. This technique enables GUV production across a broad spectrum of buffer conditions, while preventing the leakage of the encapsulated components into the oil phase. Furthermore, a simple centrifugation process is sufficient for the release of the emulsion-trapped GUVs, bypassing the need to use emulsion-destabilising chemicals. With fluorescent FNPs and transmission electron microscopy, the authors confirm that FNPs are efficiently removed, producing contaminant-free GUVs. Further experiments assessing the lateral diffusion of lipids and unilamellarity of the GUVs demonstrate that they are comparable to GUVs produced via electroformation. Finally, the ability of incorporating transmembrane proteins is demonstrated, highlighting the potential of this method for the production of GUVs for artificial cell applications.

摘要

在人工细胞的制造过程中,一个有待解决的关键挑战是复制复杂的细胞膜。在现有的模型中,巨大的单分子层囊泡(GUVs)由于其内在的分隔能力以及与真核细胞在大小和形状上的相似性而特别有趣。已经开发出许多生产 GUVs 的技术,这些技术都具有固有优势和劣势。在这里,作者展示了用于在氟油中形成 Pickering 乳液的氟化硅纳米颗粒(FNPs)可以使脂质纳米系统失稳,从而模板形成 GUVs。这种技术能够在广泛的缓冲条件下生产 GUVs,同时防止包封成分漏入油相。此外,简单的离心过程足以释放乳液捕获的 GUVs,无需使用乳液破坏化学物质。通过荧光 FNPs 和透射电子显微镜,作者证实 FNPs 被有效地去除,从而产生无污染物的 GUVs。进一步的实验评估了脂质的侧向扩散和 GUV 的单分子层性,表明它们与通过电形成生产的 GUVs相当。最后,展示了整合跨膜蛋白的能力,突出了这种方法在生产用于人工细胞应用的 GUVs 方面的潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b045/10700689/ad43e44d201e/ADVS-10-2302461-g003.jpg

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