用于确定细胞膜通透性的高占用的单细胞捕获微阵列,Lab on a Chip

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用于确定细胞膜通透性的高占用的单细胞捕获微阵列
Lab on a Chip ( IF 6.1 ) Pub Date : 2017-10-18 00:00:00 , DOI: 10.1039/c7lc00883j
Lindong Weng _{1,

2,

3,

4,

5} , Felix Ellett _{1,

2,

3,

4,

5} , Jon Edd _{1,

2,

3,

4,

5} , Keith H. K. Wong _{1,

2,

3,

4,

5} , Korkut Uygun _{1,

2,

3,

4,

5} , Daniel Irimia _{1,

2,

3,

4,

5} , Shannon L. Stott _{1,

2,

3,

4,

5} , Mehmet Toner _{1,

2,

3,

4,

5}

Affiliation

半渗透性和选择性渗透性是细胞膜的根本重要特征。膜通透性可以通过监测暴露于非等渗环境后细胞的体积变化来确定。为此目的，最近已经开发了几个微流体灌注室。但是，这些设备仅允许观察一个单个细胞或一组可能以不受控制的方式相互作用的细胞。这些设备中的一些已集成了片上温度控制，以研究膜渗透性的温度依赖性，但它们不可避免地需要复杂的制造和组装以及精细的温度和压力校准。因此，非常需要设计一种简单的单细胞捕获设备，该设备可并行监控多个单独的，各个细胞在不同温度下进行非等渗暴露。在这项研究中，我们开发了具有高陷阱占用率的单细胞无泵单层微阵列。该设备的基准性能是通过将直径为18.8μm的球形颗粒作为模型来进行的，陷阱捕获率高达86.8％，行与行之间的偏移为10–30μm。还揭示出，在每个阵列中，大于确定的临界尺寸的颗粒将在确定的横向位移模式下被捕集器排除。为了证明这种方法的实用性，我们在4、22和37°C下使用单细胞捕获装置确定大鼠肝细胞和患者源性循环肿瘤细胞（Brx-142）的膜通透性。通过脂质和水通道蛋白介导的途径。但是，Brx-142细胞的膜通透性低于大鼠肝细胞，这与水和DMSO转运的强耦合有关，但水与甘油之间的相互作用较小。此处报道的膜通透性数据为膜运输的生物物理学提供了新的见解，例如水通道蛋白表达以及水和溶质的耦合运输，以及为稀有细胞和珍贵组织生物储备的最终目标提供了必要的数据。

"点击查看英文标题和摘要"

A highly-occupied, single-cell trapping microarray for determination of cell membrane permeability

Semi- and selective permeability is a fundamentally important characteristic of the cell membrane. Membrane permeability can be determined by monitoring the volumetric change of cells following exposure to a non-isotonic environment. For this purpose, several microfluidic perfusion chambers have been developed recently. However, these devices only allow the observation of one single cell or a group of cells that may interact with one another in an uncontrolled way. Some of these devices have integrated on-chip temperature control to investigate the temperature-dependence of membrane permeability, but they inevitably require sophisticated fabrication and assembly, and delicate temperature and pressure calibration. Therefore, it is highly desirable to design a simple single-cell trapping device that allows parallel monitoring of multiple separate, individual cells subjected to non-isotonic exposure at various temperatures. In this study, we developed a pumpless, single-layer microarray with high trap occupancy of single cells. The benchmark performance of the device was conducted by targeting spherical particles of 18.8 μm in diameter as a model, yielding trap occupancy of up to 86.8% with a row-to-row shift of 10–30 μm. It was also revealed that in each array the particles larger than a corresponding critical size would be excluded by the traps in a deterministic lateral displacement mode. Demonstrating the utility of this approach, we used the single-cell trapping device to determine the membrane permeability of rat hepatocytes and patient-derived circulating tumor cells (Brx-142) at 4, 22 and 37 °C. The membrane of rat hepatocytes was found to be highly permeable to water and small molecules such as DMSO and glycerol, via both lipid- and aquaporin-mediated pathways. Brx-142 cells, however, displayed lower membrane permeability than rat hepatocytes, which was associated with strong coupling of water and DMSO transport but less interaction between water and glycerol. The membrane permeability data reported here provide new insights into the biophysics of membrane transport such as aquaporin expression and coupling transport of water and solutes, as well as providing essential data for the ultimate goal of biobanking rare cells and precious tissues.

更新日期：2017-10-25

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