| Autor: |
Dinani R; Division of Physiology, Amsterdam UMC, Vrije University; Heart Failure & Arrhythmias, Amsterdam Cardiovascular Sciences., Manders E; Division of Physiology, Amsterdam UMC, Vrije University; Heart Failure & Arrhythmias, Amsterdam Cardiovascular Sciences; CytoCypher BV., Helmes M; Division of Physiology, Amsterdam UMC, Vrije University; Heart Failure & Arrhythmias, Amsterdam Cardiovascular Sciences; CytoCypher BV., Wang L; Division of Clinical Pharmacology, Vanderbilt School of Medicine., Knollmann B; Division of Clinical Pharmacology, Vanderbilt School of Medicine., Kuster DWD; Division of Physiology, Amsterdam UMC, Vrije University; Heart Failure & Arrhythmias, Amsterdam Cardiovascular Sciences., van der Velden J; Division of Physiology, Amsterdam UMC, Vrije University; Heart Failure & Arrhythmias, Amsterdam Cardiovascular Sciences; j.vandervelden1@amsterdamumc.nl. |
| Abstrakt: |
Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) represent a powerful tool for studying mutation-mediated changes in cardiomyocyte function and defining the effects of stressors and drug interventions. In this study, it is demonstrated that this optics-based system is a powerful tool to assess the functional parameters of hiPSC-CMs in 2D. By using this platform, it is possible to perform paired measurements in a well-preserved temperature environment on different plate layouts. Moreover, this system provides researchers with instant data analysis. This paper describes a method for measuring the contractility of unmodified hiPSC-CMs. Contraction kinetics are measured at 37 °C based on pixel correlation changes relative to a reference frame taken at relaxation at a 250 Hz sampling frequency. Additionally, simultaneous measurements of intracellular calcium transients can be acquired by loading the cell with a calcium-sensitive fluorophore, such as Fura-2. Using a hyperswitch, ratiometric calcium measurements can be performed on a 50 µm diameter illumination spot, corresponding to the area of the contractility measurements. |
