Compartments for the spatially and temporally controlled assembly of biological processes are essential towards cellular life. Synthetic mimics of cellular compartments based on lipid-based protocells lack the mechanical and chemical stability to allow their manipulation into a complex and fully functional synthetic cell. Here, we present a high-throughput microfluidic method to generate stable, defined sized liposomes termed ‘droplet-stabilized giant unilamellar vesicles (dsGUVs)’. The enhanced stability of dsGUVs enables the sequential loading of these compartments with biomolecules, namely purified transmembrane and cytoskeleton proteins by microfluidic pico-injection technology. This constitutes an experimental demonstration of a successful bottom-up assembly of a compartment with contents that would not self-assemble to full functionality when simply mixed together. Following assembly, the stabilizing oil phase and droplet shells are removed to release functional self-supporting protocells to an aqueous phase, enabling them to interact with physiologically relevant matrices.

在空间和时间上控制生物过程装配的隔室对于细胞生命至关重要。基于脂质的原始细胞的细胞区室的合成模拟物缺乏机械和化学稳定性，无法将其操纵为复杂且功能齐全的合成细胞。在这里，我们提出了一种高通量的微流体方法，以生成稳定的，确定大小的脂质体，称为“液滴稳定的巨型单层囊泡（dsGUVs）”。dsGUV的增强的稳定性使这些小室能够通过微流体皮层注射技术依次装载生物分子，即纯化的跨膜蛋白和细胞骨架蛋白。这构成了一个成功的自底向上组装式隔间的实验性演示，该隔间的内容物在简单地混合在一起时不会自组装成完整的功能。组装后，将稳定的油相和液滴壳去除，以将功能性的自支撑原细胞释放到水相中，从而使其与生理相关的基质相互作用。