High-Throughput Phenotyping of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes and Neurons Using Electric Field Stimulation and High-Speed Fluorescence Imaging

被引:7
作者
Daily, Neil J. [1 ]
Du, Zhong-Wei [2 ]
Wakatsuki, Tetsuro [1 ]
机构
[1] InvivoSciences Inc, 510 Charmany Dr,Suite 265, Madison, WI 53719 USA
[2] BrainXell Inc, Madison, WI USA
关键词
Cal-520; calcium; cardiomyocyte; electric field stimulation; FLIPR Calcium 6; induced pluripotent stem cell; PRECLINICAL CARDIAC ELECTROPHYSIOLOGY; SYNAPTIC-TRANSMISSION; ACTION-POTENTIALS; MYOCYTE CULTURES; MOTOR-NEURONS; IN-VITRO; ALTERNANS; MECHANISMS; HYPEREXCITABILITY; PROLONGATION;
D O I
10.1089/adt.2017.781
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Electrophysiology of excitable cells, including muscle cells and neurons, has been measured by making direct contact with a single cell using a micropipette electrode. To increase the assay throughput, optical devices such as microscopes and microplate readers have been used to analyze electrophysiology of multiple cells. We have established a high-throughput (HTP) analysis of action potentials (APs) in highly enriched motor neurons and cardiomyocytes (CMs) that are differentiated from human induced pluripotent stem cells (iPSCs). A multichannel electric field stimulation (EFS) device enabled the ability to electrically stimulate cells and measure dynamic changes in APs of excitable cells ultra-rapidly (> 100 data points per second) by imaging entire 96-well plates. We found that the activities of both neurons and CMs and their response to EFS and chemicals are readily discerned by our fluorescence imaging-based HTP phenotyping assay. The latest generation of calcium (Ca2+) indicator dyes, FLIPR Calcium 6 and Cal-520, with the HTP device enables physiological analysis of human iPSC-derived samples highlighting its potential application for understanding disease mechanisms and discovering new therapeutic treatments.
引用
收藏
页码:178 / 188
页数:11
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