Department of Biomedical Engineering, The George Washington University, Washington, DC, USA.
Lab Chip. 2020 Nov 7;20(21):4031-4042. doi: 10.1039/d0lc00615g. Epub 2020 Sep 30.
This work demonstrates a novel high-throughput (HT) microfluidics-enabled uninterrupted perfusion system (HT-μUPS) and validates its use with chronic all-optical electrophysiology in human excitable cells. HT-μUPS consists of a soft multichannel microfluidic plate cover which could button on a commercial HT 96-well plate. Herein, we demonstrate the manufacturing process of the system and its usages in acute and chronic all-optical electrophysiological studies of human induced pluripotent stem-cell-derived cardiomyocytes (iPSC-CM) and engineered excitable (spiking HEK) cells. HT-μUPS perfusion maintained functional voltage and calcium responses in iPSC-CM and spiking HEK cells under spontaneous conditions and under optogenetic pacing. Long-term culture with HT-μUPS improved cell viability and optogenetically-tracked calcium responses in spiking HEK cells. The simplicity of this design and its compatibility with HT all-optical electrophysiology can empower cell-based assays for personalized medicine using patient-derived cells.
这项工作展示了一种新颖的高通量（HT）微流控不间断灌流系统（HT-μUPS），并通过人类可兴奋细胞的慢性全光学电生理学验证了其用途。HT-μUPS 由一个软多通道微流控板盖组成，可以扣在商业 HT96 孔板上。本文介绍了该系统的制造工艺及其在急性和慢性全光学电生理学研究中的应用，包括人诱导多能干细胞衍生的心肌细胞（iPSC-CM）和工程可兴奋（起搏 HEK）细胞。HT-μUPS 灌流在自发条件下和光遗传起搏下维持了 iPSC-CM 和起搏 HEK 细胞的功能电压和钙反应。HT-μUPS 的长期培养提高了起搏 HEK 细胞的细胞活力和光遗传追踪的钙反应。该设计的简单性及其与 HT 全光学电生理学的兼容性，可以为使用患者来源细胞的个性化医学提供基于细胞的检测。
