Huang Xiaowen, Yue Wanqing, Liu Dandan, Yue Jianbo, Li Jiaqian, Sun Dong, Yang Mengsu, Wang Zuankai
Department of Mechanical and Biomedical Engineering,City University of Hong Kong, Hong Kong, China.
Department of Biomedical Sciences,City University of Hong Kong, Hong Kong, China.
Sci Rep. 2016 Mar 23;6:23591. doi: 10.1038/srep23591.
The profiling of physiological response of cells to external stimuli at the single cell level is of importance. Traditional approaches to study cell responses are often limited by ensemble measurement, which is challenging to reveal the complex single cell behaviors under a dynamic environment. Here we report the development of a simple microfluidic device to investigate intracellular calcium response to dynamic hypertonic conditions at the single cell level in real-time. Interestingly, a dramatic elevation in the intracellular calcium signaling is found in both suspension cells (human leukemic cell line, HL-60) and adherent cells (lung cancer cell line, A549), which is ascribed to the exposure of cells to the hydrodynamic stress. We also demonstrate that the calcium response exhibits distinct single cell heterogeneity as well as cell-type-dependent responses to the same stimuli. Our study opens up a new tool for tracking cellular activity at the single cell level in real time for high throughput drug screening.
在单细胞水平上分析细胞对外部刺激的生理反应具有重要意义。传统的研究细胞反应的方法通常受到整体测量的限制，在动态环境下揭示复杂的单细胞行为具有挑战性。在此，我们报告了一种简单微流控装置的开发，用于实时研究单细胞水平下细胞内钙对动态高渗条件的反应。有趣的是，在悬浮细胞（人白血病细胞系HL-60）和贴壁细胞（肺癌细胞系A549）中均发现细胞内钙信号显著升高，这归因于细胞受到流体动力应力的作用。我们还证明，钙反应表现出明显的单细胞异质性以及对相同刺激的细胞类型依赖性反应。我们的研究为在单细胞水平实时跟踪细胞活动以进行高通量药物筛选开辟了一种新工具。
