Contractions of Human-iPSC-derived Cardiomyocyte Syncytia Measured with a Ca-sensitive Fluorescent Dye in Temperature-controlled 384-well Plates

Abstract

Spontaneously contracting syncytia of cardiomyocytes derived from human-induced pluripotent stem cells (hiPSC-CM) are a useful model of human cardiac physiology and pharmacology. Various methods have been proposed to record this spontaneous activity and to evaluate drug effects, but many of these methods suffer from limited throughput and/or physiological relevance. We developed a high-throughput screening system to quantify the effects of exogenous compounds on hiPSC-CM's beating frequency, using a Ca-sensitive fluorescent dye and a temperature-controlled imaging multi-well plate reader. We describe how to prepare the cell plates and the compound plates and how to run the automated assay to achieve high sensitivity and reproducibility. We also describe how to transform and analyze the fluorescence data to provide reliable measures of drug effects on spontaneous rhythm. This assay can be used in drug discovery programs to guide chemical optimization away from, or toward, compounds affecting human cardiac function.

Creating cell-specific computational models of stem cell-derived cardiomyocytes using optical experiments

“…To simulate a heterogeneous set of iPSC-CM cell lines, a "population" of Kernik model cells was created by drawing multiplier factors for each maximal conductance parameter from a log-normal distribution with mean multiplier = 1, spread = 0.2. This population was then simulated under physiological, unperturbed conditions (Supplementary Table S3, protocol 19), and filtered for cells which showed a spontaneous beating frequency between 0.3 Hz and 1.0 Hz, nearly matching the automaticity rate range for human iPSC-CMs reported in [50]. Out of the remaining model cells, 4 were randomly selected for simulation under the 19 different conditions, to generate the final in silico dataset.…”