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@proceedings{2417402, author = {Švecová, Olga and Král, Martin and Zelenák, Štefan and Pacherník, Jiří and Bárta, Tomáš and Lietava, Samuel and Synková, Iva and Zídková, Jana and Novotný, Tomáš and Bébarová, Markéta}, booktitle = {15th New Frontiers in Basic Cardiovascular Research: A France-New EU Members Symposium}, keywords = {calcium transient; hiPSC-derived cardiomyocytes}, language = {eng}, title = {Analysis of calcium transients in cardiomyocytes derived from hiPSCs: the variant p. Y4734C in RYR2 vs. unrelated healthy controls}, year = {2024} }
TY - CONF ID - 2417402 AU - Švecová, Olga - Král, Martin - Zelenák, Štefan - Pacherník, Jiří - Bárta, Tomáš - Lietava, Samuel - Synková, Iva - Zídková, Jana - Novotný, Tomáš - Bébarová, Markéta PY - 2024 TI - Analysis of calcium transients in cardiomyocytes derived from hiPSCs: the variant p. Y4734C in RYR2 vs. unrelated healthy controls KW - calcium transient KW - hiPSC-derived cardiomyocytes N2 - Introduction: the regulation of intracellular calcium levels is crucial for excitation-contraction coupling. The release of calcium into the intracellular space is controlled by the ryanodine receptor type 2 (RYR2) located on the sarcoplasmic reticulum. Dysfunction of RYR2 is involved in the pathogenesis of inherited and non-inherited diseases such as cardiac arrhythmias, ventricular fibrillation, ventricular tachycardia, etc. The variant p. Y4734C in RYR2 was found in a patient with idiopathic ventricular fibrillation without structural changes in the heart and signs of arrhythmia on clinical examination. Preliminary data show calcium transient of patient-specific cardiomyocytes derived from human-induced pluripotent stem cells (hiPSC-CM). Methods: calcium transients of hiPSC-CM (Y4734C) and hiPSC-CM unrelated healthy controls (WT) were measured using the Myocyte Calcium and Contractility System (IonOptix LLC). Cell clusters were loaded with Fura-2 (Molecular Probes, Invitrogen) at a final concentration of 1 mM. Cells were incubated for 15 min in Tyrode solution with 1 μmol/L Fura-2-am at 37 °C and then washed repeatedly with Tyrode solution followed by incubation for 10 min in Tyrode solution at 37 °C. Measurements were performed in Tyrode solution at 37 ± 0.5 °C. Cells were not stimulated. Analysis of calcium transients was performed using CytoSolver software (IonOptix LLC). Results: calcium transient parameters and frequency of Y4734C and WT were evaluated. Time to peak and Time constant were significantly longer in Y4734C (0,24±xx and 0,20±xx s, respectively; n=8; P˂0.004 and P˂0.048) than in WT (0,09±xx and 0,11±xx s, respectively; n=4). A nonsignificant change was observed in the amplitude of the calcium transient, Y4734C (0,22±xx; n=8) and WT (0,15±xx; n=4). Cell clusters with the variant Y4734C have higher frequency then WT (40±xx and 28±xx b/min, respectively). Conclusions: the preliminary data showed delayed release of calcium from the sarcoplasmic reticulum in cells with the variant Y4734C and prolonged reabsorption of calcium back into the sarcoplasmic reticulum. ER -
ŠVECOVÁ, Olga, Martin KRÁL, Štefan ZELENÁK, Jiří PACHERNÍK, Tomáš BÁRTA, Samuel LIETAVA, Iva SYNKOVÁ, Jana ZÍDKOVÁ, Tomáš NOVOTNÝ a Markéta BÉBAROVÁ. Analysis of calcium transients in cardiomyocytes derived from hiPSCs: the variant p. Y4734C in RYR2 vs. unrelated healthy controls. In \textit{15th New Frontiers in Basic Cardiovascular Research: A France-New EU Members Symposium}. 2024.
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