Contractions of Human-iPSC-derived Cardiomyocyte Syncytia Measured with a Ca-sensitive Fluorescent Dye in Temperature-controlled 384-well Plates
| Autor: | Camille Forny, Romain Sube, Eric A. Ertel |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
General Chemical Engineering Phenotypic screening Induced Pluripotent Stem Cells cardiomyocytes Issue 140 rhythm iPS cell General Biochemistry Genetics and Molecular Biology 03 medical and health sciences safety pharmacology Myocyte Myocytes Cardiac Induced pluripotent stem cell Fluorescent Dyes Syncytium General Immunology and Microbiology Drug discovery Chemistry General Neuroscience Safety pharmacology phenotypic screening Temperature Fluorescence Phenotype 030104 developmental biology Biophysics Medicine pharmacology Plate reader |
| Zdroj: | Journal of Visualized Experiments : JoVE |
| ISSN: | 1940-087X |
| DOI: | 10.3791/58290 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|