Microfluidic production of giant lipid vesicles presents a paradigm-shift in the development of artificial cells. While production is high-throughput and the lipid vesicles are mono-disperse compared to bulk methods, current technologies rely heavily on the addition of additives such as surfactants, glycerol and even ethanol. Here we present a microfluidic method for producing biomimetic surfactant-free and additive-free giant unilamellar vesicles. The versatile design allows for the production of vesicle sizes ranging anywhere from ~10 to 130 µm with either neutral or charged lipids, and in physiological buffer conditions. Purity, functionality, and stability of the membranes are validated by lipid diffusion, protein incorporation, and leakage assays. Usability as artificial cells is demonstrated by increasing their complexity, i.e., by encapsulating plasmids, smaller liposomes, mammalian cells, and microspheres. This robust method capable of creating truly biomimetic artificial cells in high-throughput will prove valuable for bottom-up synthetic biology and the understanding of membrane function.

巨型脂质囊泡的微流控生产代表了人工细胞发展的范式转变。虽然生产是高通量的，并且与批量方法相比，脂质囊泡是单分散的，但目前的技术在很大程度上依赖于添加表面活性剂、甘油甚至乙醇等添加剂。在这里，我们提出了一种用于生产无仿生表面活性剂和无添加剂的巨型单层囊泡的微流体方法。多功能设计允许在生理缓冲条件下使用中性或带电脂质生产大小从 ~10 到 130 µm 不等的囊泡。膜的纯度、功能和稳定性通过脂质扩散、蛋白质掺入和渗漏测定进行验证。通过增加它们的复杂性来证明作为人造细胞的可用性，即 通过封装质粒、较小的脂质体、哺乳动物细胞和微球。这种能够以高通量创建真正的仿生人造细胞的稳健方法将证明对自下而上的合成生物学和对膜功能的理解很有价值。