Monitoring Human-Induced Pluripotent Stem Cell-Derived Cardiomyocytes with Genetically Encoded Calcium and Voltage Fluorescent Reporters

Abstract

SummaryThe advent of the human-induced pluripotent stem cell (hiPSC) technology has transformed biomedical research, providing new tools for human disease modeling, drug development, and regenerative medicine. To fulfill its unique potential in the cardiovascular field, efficient methods should be developed for high-resolution, large-scale, long-term, and serial functional cellular phenotyping of hiPSC-derived cardiomyocytes (hiPSC-CMs). To achieve this goal, we combined the hiPSC technology with genetically encoded voltage (ArcLight) and calcium (GCaMP5G) fluorescent indicators. Expression of ArcLight and GCaMP5G in hiPSC-CMs permitted to reliably follow changes in transmembrane potential and intracellular calcium levels, respectively. This allowed monitoring short- and long-term changes in action-potential and calcium-handling properties and the development of arrhythmias in response to several pharmaceutical agents and in hiPSC-CMs derived from patients with different inherited arrhythmogenic syndromes. Combining genetically encoded fluorescent reporters with hiPSC-CMs may bring a unique value to the study of inherited disorders, developmental biology, and drug development and testing.

Patient-Specific Drug Screening Using a Human Induced Pluripotent Stem Cell Model of Catecholaminergic Polymorphic Ventricular Tachycardia Type 2

“…Subsequently, many hiPSC-CMs models of both CPVT types [20][21][22][23][24][25][26][27][28][29] were shown to recapitulate the clinical phenotype in the culture-dish by displaying abnormal Ca 2+ handling and arrhythmias. Here, we focused on CPVT2 and found, similar to previous reports, 24,25,29 21,30 We recently investigated this phenomenon in a CPVT1-hiPSC-CMs model demonstrating a reduced SOICR threshold in the affected cells. 21 Here, we demonstrated the ability to model SOICR also in the CPVT2-hiPSC-CMs and revealed that the D307H-CASQ2 may also reduce SOICR threshold in the affected cardiomyocytes.…”

“…7,8 In the cardiac field, hiPSCs were established from healthy individuals and from patients inflicted with acquired 9 and several types of inherited cardiac disorders. 10,11 Patient-specific hiPSC-derived cardiomyocyte (hiPSC-CM) models of different inherited arrhythmogenic syndromes (including the long-QT, [12][13][14][15] Brugada, 16 and sodium channel overlap 17 syndromes; arrhythmogenic right ventricular cardiomyopathy 18,19 ; and both types of CPVT [20][21][22][23][24][25][26][27][28][29] ) were…”

“…We used previously well-established healthy control and CPVT2 hiPSC lines, 29 the latter derived from a 19-year-old male patient with CPVT2 because of the D307H-CASQ2 mutation. 29 Dermal fibroblasts were used for the creation of the hiPSC lines.…”

“…29 Dermal fibroblasts were used for the creation of the hiPSC lines. Cardiomyocyte differentiation was induced using the monolayer differentiation system.…”

“…Cardiomyocyte differentiation was induced using the monolayer differentiation system. 29 In brief, hiPSC colonies were propagated on Matrigel using mTeSR-1 (Stem-Cell Technologies). Cardiomyocyte induction was performed using a differentiation medium containing RPMI-1640, 2%-B27 supplement minus insulin (Life Technologies), 1% penicillin/streptomycin, and 6 μmol/L CHIR99021 (Stemgent) for 2 days.…”