A Controllable, Centrifugal-Based Hydrodynamic Microfluidic Chip for Cell-Pairing and Studying Long-Term Communications between Single Cells
A Controllable, Centrifugal-Based Hydrodynamic Microfluidic Chip for Cell-Pairing and Studying Long-Term Communications between Single Cells
复制标题
实验设计微流控芯片上干细胞分化因子组合的优化
DOI：
10.1021/acs.analchem.9b04370
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发表时间：
2019-12-17
影响因子：
7.4
通讯作者：
Wu, Hongkai
中科院分区：
文献类型：
--
作者：
Li, Lijun;Wang, Huirong;Wu, Hongkai
在明确定义的微环境中，研究单个细胞之间的细胞相互作用对于理解特定的细胞间通信和相互作用至关重要。然而，目前大多数多细胞系统的研究往往被其他复杂的相互作用所淹没。基于微流控芯片的细胞配对为简化研究提供了一种潜在的策略。在这里，我们报告了一种强大而直接的方法，依赖于流体动力学单细胞捕获和单个细胞离心辅助重新定位的组合，通常可以应用于各种细胞类型的细胞配对和研究单细胞水平的细胞相互作用。这种微流控芯片操作简单，易于控制，只需要两个操作步骤-用流体动力学陷阱捕获单个细胞，随后通过离心重新定位捕获细胞。利用这种微流控芯片，我们证明了同型细胞配对、异型细胞配对和长期细胞共培养，与以前的细胞配对方法相比，它们表现出更好或相当的性能。其单细胞捕获效率和细胞配对效率分别接近74%和20%。作为概念验证，我们将单个dHL-60细胞和HeLa细胞（HeLa- il8、HeLa- il10和野生型HeLa细胞）在多个细胞室中配对。HeLa-IL8和HeLa-ILIO均采用光诱导基因表达系统，在蓝光照射下分别分泌白细胞介素-8和白细胞介素-10。我们发现这三种HeLa细胞系对dHL-60细胞迁移的影响非常不同。该平台展示了其在细胞间通讯（旁分泌）研究中的潜在应用，并且可以扩展到捕获更复杂生物系统中的三个或更多单个细胞。
Investigation of cell-cell interactions between individual cells in a well-defined microenvironment is critical for the understanding of specific intercellular communications and interactions. However, most current studies in multicellular systems are often overwhelmed by additional complicated interactions. Cell-pairing based on a microfluidic chip provides a potential strategy to simplify the studies. Here, we report a robust and straightforward method, relying on a combination of hydrodynamic single-cell capture and centrifugation-assisted relocation of individual cells, which can be applied, in general, to various cell types for cell-pairing and studying cell interactions at the single-cell level. This microfluidic chip is simple to operate and easily controlled, which requires only two operational steps-capturing individual cells with hydrodynamic traps and subsequently relocating the capture cells by centrifugation. With this microfluidic chip, we demonstrated homotypic cell-pairing, heterotypic cell-pairing, and long-term cell coculture, which exhibited better or comparable performance compared with previous cell-pairing methods. Its single-cell trapping and cell-pairing efficiencies are similar to 74% and similar to 20%, respectively. As a proof of concept, we paired individual dHL-60 cells and HeLa cells (HeLa-IL8, HeLa-IL10, and wild-type HeLa cells) in multiple cell chambers. The HeLa-IL8 and HeLa-ILIO, both engineered with a light-induced gene expression system, can secret interleukin-8 and interleukin-10, respectively, under blue light illumination. We found that these three HeLa cell lines have very different influences on the migration of dHL-60 cells. This platform demonstrates its potential applications in studies of intercellular communication (paracrine), and it can be extended to trap three or more individual cells for more complex biological systems.