Department of Chemistry , The Hong Kong University of Science and Technology , Clear Water Bay, Kowloon , Hong Kong , China.
Department of Biology, Guangdong Provincial Key Laboratory of Cell Microenvironment and Disease Research, Shenzhen Key Laboratory of Cell Microenvironment , Southern University of Science and Technology , Shenzhen 518055 , China.
Anal Chem. 2019 Dec 17;91(24):15908-15914. doi: 10.1021/acs.analchem.9b04370. Epub 2019 Dec 4.
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 ∼74% and ∼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-IL10, 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.
在明确的微环境中研究单个细胞之间的细胞-细胞相互作用对于理解特定的细胞间通讯和相互作用至关重要。然而，大多数当前的多细胞系统研究往往受到额外的复杂相互作用的困扰。基于微流控芯片的细胞配对提供了一种简化研究的潜在策略。在这里，我们报告了一种稳健且简单的方法，该方法依赖于流体动力学单细胞捕获和离心辅助单个细胞重定位的结合，该方法通常可应用于各种细胞类型，用于单细胞水平的细胞配对和细胞相互作用研究。这种微流控芯片操作简单，易于控制，仅需两个操作步骤-使用流体动力阱捕获单个细胞，然后通过离心重新定位捕获的细胞。使用这种微流控芯片，我们展示了同型细胞配对、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，其表现更好或相当。其单细胞捕获和细胞配对效率分别约为 74%和 20%。作为概念验证，我们在多个细胞室中配对了单个 dHL-60 细胞和 HeLa 细胞（HeLa-IL8、HeLa-IL10 和野生型 HeLa 细胞）。这两种 HeLa 细胞系均通过光诱导基因表达系统构建，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 细胞的迁移有非常不同的影响。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三个或更多个单个细胞以用于更复杂的生物系统。
