Droplet Array-Based Platform for Parallel Optical Analysis of Dynamic Extracellular Vesicle Secretion from Single Cells
Droplet Array-Based Platform for Parallel Optical Analysis of Dynamic Extracellular Vesicle Secretion from Single Cells
复制标题
基于液滴阵列的单细胞动态细胞外囊泡分泌并行光学分析平台
DOI：
10.1021/acs.analchem.2c01609
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发表时间：
2022
影响因子：
7.4
通讯作者：
Ota Sadao
中科院分区：
文献类型：
--
作者：
Hattori Kazuki;Goda Yuki;Yamashita Minato;Yoshioka Yusuke;Kojima Ryosuke;Ota Sadao
细胞外囊泡（extracellular vesicles，EVs）是细胞间重要的通讯工具，但由于缺乏精确的分析方法，其异质性分泌的调控机制尚不清楚。监测来自单独分离的细胞的分泌的动力学是至关重要的，因为在批量分析中，分泌活性可以被细胞-细胞相互作用干扰，并且细胞群体很少以幅度或持续时间同步的方式进行分泌。虽然已经采用了各种微流体技术来评估单细胞衍生EV的丰度，但没有一种技术可以长时间持续跟踪它们的分泌动力学。在这里，我们开发了一种基于液滴阵列的方法，该方法允许我们光学定量>300个单细胞的EV分泌动力学，每2小时持续36小时，这涵盖了许多细胞类型的细胞倍增时间。实验结果清楚地显示了单细胞EV分泌的高度异质性，并表明细胞分裂促进EV分泌，显示了该平台用于发现EV调节机制的有用性。
Extracellular vesicles (EVs) are essential intercellular communication tools, but the regulatory mechanisms governing heterogeneous EV secretion are still unclear due to the lack of methods for precise analysis. Monitoring the dynamics of secretion from individually isolated cells is crucial because in bulk analysis, secretion activity can be perturbed by cell–cell interactions, and a cell population rarely performs secretion in a magnitude- or duration-synchronized manner. Although various microfluidic techniques have been adopted to evaluate the abundance of single-cell-derived EVs, none can track their secretion dynamics continually for extended periods. Here, we have developed a droplet array-based method that allowed us to optically quantify the EV secretion dynamics of >300 single cells every 2 h for 36 h, which covers the cell doubling time of many cell types. The experimental results clearly show the highly heterogeneous nature of single-cell EV secretion and suggest that cell division facilitates EV secretion, showing the usefulness of this platform for discovering EV regulation machinery.