title: High throughput measurement of Ca²⁺ dynamics for drug risk assessment in human stem cell-derived cardiomyocytes by kinetic image cytometry.
abstract: Current methods to measure physiological properties of cardiomyocytes and predict fatal arrhythmias that can cause sudden death, such as Torsade de Pointes, lack either the automation and throughput needed for early-stage drug discovery and/or have poor predictive value. To increase throughput and predictive power of in vitro assays, we developed kinetic imaging cytometry (KIC) for automated cell-by-cell analyses via intracellular fluorescence Ca²⁺ indicators. The KIC instrument simultaneously records and analyzes intracellular calcium concentration [Ca²⁺](i) at 30-ms resolution from hundreds of individual cells/well of 96-well plates in seconds, providing kinetic details not previously possible with well averaging technologies such as plate readers. Analyses of human embryonic stem cell and induced pluripotent stem cell-derived cardiomyocytes revealed effects of known cardiotoxic and arrhythmogenic drugs on kinetic parameters of Ca²⁺ dynamics, suggesting that KIC will aid in the assessment of cardiotoxic risk and in the elucidation of pathogenic mechanisms of heart disease associated with drugs treatment and/or genetic background.
authors.affiliation: Sanford-Burnham Medical Research Institute, 10901N. Torrey Pines Road, La Jolla, CA 92037, USA.
authors.forename: Fabio
authors.initials: F
authors.lastname: Cerignoli
authors.forename: David
authors.initials: D
authors.lastname: Charlot
authors.forename: Ross
authors.initials: R
authors.lastname: Whittaker
authors.forename: Randy
authors.initials: R
authors.lastname: Ingermanson
authors.forename: Piyush
authors.initials: P
authors.lastname: Gehalot
authors.forename: Alex
authors.initials: A
authors.lastname: Savchenko
authors.forename: David J
authors.initials: DJ
authors.lastname: Gallacher
authors.forename: Rob
authors.initials: R
authors.lastname: Towart
authors.forename: Jeffrey H
authors.initials: JH
authors.lastname: Price
authors.forename: Patrick M
authors.initials: PM
authors.lastname: McDonough
authors.forename: Mark
authors.initials: M
authors.lastname: Mercola
mesh_terms: D000818:Animals; D001145:Arrhythmias, Cardiac / Q000139:chemically induced; D001331:Automation; D002118:Calcium / Q000378:metabolism*; D055808:Drug Discovery / Q000379:methods; D064420:Drug-Related Side Effects and Adverse Reactions; D053595:Embryonic Stem Cells / Q000378:metabolism; D005453:Fluorescence; D006331:Heart Diseases / Q000139:chemically induced / Q000235:genetics / Q000503:physiopathology; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D019044:Image Cytometry / Q000379:methods; D032383:Myocytes, Cardiac / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D039904:Pluripotent Stem Cells / Q000378:metabolism; D011237:Predictive Value of Tests; D051381:Rats; D018570:Risk Assessment / Q000379:methods
chemical_list: D002118:Calcium