Department of Bionanoscience, Kavli Institute of Nanoscience Delft , Delft University of Technology , Van der Maasweg 9 , 2629 HZ Delft , The Netherlands.
ACS Nano. 2018 Mar 27;12(3):2560-2568. doi: 10.1021/acsnano.7b08411. Epub 2018 Feb 22.
Liposomes, self-assembled vesicles with a lipid-bilayer boundary similar to cell membranes, are extensively used in both fundamental and applied sciences. Manipulation of their physical properties, such as growth and division, may significantly expand their use as model systems in cellular and synthetic biology. Several approaches have been explored to controllably divide liposomes, such as shape transformation through temperature cycling, incorporation of additional lipids, and the encapsulation of protein division machinery. However, so far, these methods lacked control, exhibited low efficiency, and yielded asymmetric division in terms of volume or lipid composition. Here, we present a microfluidics-based strategy to realize mechanical division of cell-sized (∼6 μm) liposomes. We use octanol-assisted liposome assembly (OLA) to produce liposomes on chip, which are subsequently flowed against the sharp edge of a wedge-shaped splitter. Upon encountering such a Y-shaped bifurcation, the liposomes are deformed and, remarkably, are able to divide into two stable daughter liposomes in just a few milliseconds. The probability of successful division is found to critically depend on the surface area-to-volume ratio of the mother liposome, which can be tuned through osmotic pressure, and to strongly correlate to the mother liposome size for given microchannel dimensions. The division process is highly symmetric (∼3% size variation between the daughter liposomes) and is accompanied by a low leakage. This mechanical division of liposomes may constitute a valuable step to establish a growth-division cycle of synthetic cells.
脂质体是一种具有类似细胞膜双层边界的自组装囊泡,广泛应用于基础科学和应用科学。操纵其物理性质,如生长和分裂,可能会极大地扩展它们作为细胞和合成生物学模型系统的用途。已经探索了几种方法来控制脂质体的分裂,例如通过温度循环进行形状转换、掺入额外的脂质以及封装蛋白分裂机制。然而,到目前为止,这些方法缺乏控制,效率低下,并且在体积或脂质组成方面表现出不对称分裂。在这里,我们提出了一种基于微流控的策略,以实现细胞大小(约 6μm)脂质体的机械分裂。我们使用辛醇辅助脂质体组装(OLA)在芯片上制备脂质体,然后将其与楔形分裂器的锐利边缘相对流动。当遇到这样的 Y 形分叉时,脂质体被变形,并且令人惊讶的是,它们能够在几毫秒内分裂成两个稳定的子脂质体。成功分裂的概率被发现严重依赖于母脂质体的表面积与体积比,这可以通过渗透压进行调节,并且与给定微通道尺寸的母脂质体大小强烈相关。分裂过程高度对称(子脂质体之间的大小变化约为 3%),并且泄漏量低。这种脂质体的机械分裂可能是建立合成细胞生长-分裂周期的重要步骤。