Jang Huisoo, Hu Peichi C, Jung Sungho, Kim Won Young, Kim Sun Min, Malmstadt Noah, Jeon Tae-Joon
Department of Biological Engineering, Inha University, Incheon, Republic of Korea; Biohybrid Systems Research Center, Inha University, Incheon, Republic of Korea.
Biotechnol J. 2013 Nov;8(11):1341-6. doi: 10.1002/biot.201200388.
Vesosomes - hierarchical assemblies consisting of membrane-bound vesicles of various scales - are potentially powerful models of cellular compartmentalization. Current methods of vesosome fabrication are labor intensive, and offer little control over the size and uniformity of the final product. In this article, we report the development of an automated vesosome formation platform using a microfluidic device and a continuous flow microcentrifuge. In the microfluidic device, water-in-oil droplets containing nanoscale vesicles in the water phase were formed using T-junction geometry, in which a lipid monolayer is formed at the oil/water interface. These water-in-oil droplets were then immediately transferred to the continuous flow microcentrifuge. When a water-in-oil droplet passed through a second lipid monolayer formed in the continuous flow microcentrifuge, a bilayer-encapsulated vesosome was created, which contained all of the contents of the aqueous phase encapsulated within the vesosome. Encapsulation of nanoscale liposomes within the outer vesosome membrane was confirmed by fluorescence microscopy. Laser diffraction analysis showed that the vesosomes we fabricated were uniform (coefficient of variation of 0.029). The yield of the continuous flow microcentrifuge is high, with over 60% of impinging water droplets being converted to vesosomes. Our system provides a fully automatable route for the generation of vesosomes encapsulating arbitrary contents. The method employed in this work is simple and can be readily applied to a variety of systems, providing a facile platform for fabricating multicomponent carriers and model cells.
囊泡体——由各种尺度的膜结合囊泡组成的分层组装体——可能是细胞区室化的强大模型。目前的囊泡体制备方法劳动强度大，且对最终产品的大小和均匀性控制有限。在本文中，我们报告了一种使用微流控装置和连续流微量离心机的自动化囊泡形成平台的开发。在微流控装置中，利用T型结几何结构形成了水相含有纳米级囊泡的油包水液滴，其中在油/水界面形成了脂质单层。然后将这些油包水液滴立即转移到连续流微量离心机中。当一个油包水液滴通过在连续流微量离心机中形成的第二个脂质单层时，就形成了一个双层包裹的囊泡体，其中包含包裹在囊泡体内的水相的所有内容物。通过荧光显微镜证实了纳米级脂质体被包裹在外囊泡膜内。激光衍射分析表明，我们制备的囊泡体是均匀的（变异系数为0.029）。连续流微量离心机的产率很高，超过60%的撞击水滴被转化为囊泡体。我们的系统为生成包裹任意内容物的囊泡体提供了一条完全自动化的途径。这项工作中采用的方法很简单，并且可以很容易地应用于各种系统，为制造多组分载体和模型细胞提供了一个简便的平台。
