论文标题
使用微流体微孔抽吸芯片的多细胞球体的高通量机械化表型
High-throughput mechanophenotyping of multicellular spheroids using a microfluidic micropipette aspiration chip
论文作者
论文摘要
细胞球体是模仿生物组织的拥挤微环境的体外多细胞模型系统。它们的机械表征可以为单细胞力学和细胞 - 细胞相互作用如何控制组织力学和自组织的宝贵见解。但是,大多数测量技术一次仅一次探测一个球体,需要专门的设备并且难以处理。在这里,我们开发了一种微流体芯片,该芯片遵循玻璃毛细管微孔抽吸的概念,以便以易于处理的高通量方式量化球体的粘弹性行为。球体通过轻柔的流动将球形装在平行口袋中,然后使用静水压力将球体舌吸入相邻的抽吸通道中。每个实验后,通过逆转压力,可以轻松从芯片中移除球体,并可以注入新的球体。具有均匀抽吸压力的多个口袋的存在,结合了进行连续实验的易感性,每天可以产生数十几个球体的吞吐量。我们证明,在以不同的抽吸压力下工作时,芯片可提供准确的变形数据。最后,我们测量由不同细胞系制成的球体的粘弹性特性,并显示了使用已建立的实验技术与以前的研究一致的。总而言之,我们的芯片提供了一种高通量的方法来测量细胞球体的粘弹性变形行为,以便进行机械型不同的组织类型,并检查细胞 - 内膜特性与整体组织行为之间的联系。
Cell spheroids are in vitro multicellular model systems that mimic the crowded micro-environment of biological tissues. Their mechanical characterization can provide valuable insights in how single-cell mechanics and cell-cell interactions control tissue mechanics and self-organization. However, most measurement techniques are limited to probing one spheroid at a time, require specialized equipment and are difficult to handle. Here, we developed a microfluidic chip that follows the concept of glass capillary micropipette aspiration in order to quantify the viscoelastic behavior of spheroids in an easy- to-handle, high-throughput manner. Spheroids are loaded in parallel pockets via a gentle flow, after which spheroid tongues are aspirated into adjacent aspiration channels using hydrostatic pressure. After each experiment, the spheroids are easily removed from the chip by reversing the pressure and new spheroids can be injected. The presence of multiple pockets with a uniform aspiration pressure, combined with the ease to conduct successive experiments, allows for a high throughput of tens of spheroids per day. We demonstrate that the chip provides accurate deformation data when working at different aspiration pressures. Lastly, we measure the viscoelastic properties of spheroids made of different cell lines and show how these are consistent with previous studies using established experimental techniques. In summary, our chip provides a high-throughput way to measure the viscoelastic deformation behavior of cell spheroids, in order to mechanophenotype different tissue types and examine the link between cell-intrinsic properties and overall tissue behavior.