TY - JOUR
T1 - Real-Time Measurements of Calcium and Contractility Parameters in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes
AU - Dinani, Rafeeh
AU - Manders, Emmy
AU - Helmes, Michiel
AU - Wang, Lili
AU - Knollmann, Bjorn
AU - Kuster, Diederik W. D.
AU - van der Velden, Jolanda
N1 - Funding Information: This research has been funded in part by Eurostars grant Estars2 113937 CARDIOMYO (EM & DK), and NWO-VICI grant 91818602 (JV). Publisher Copyright: © 2023 JoVE Journal of Visualized Experiments.
PY - 2023
Y1 - 2023
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85162004723&partnerID=8YFLogxK
U2 - https://doi.org/10.3791/65326
DO - https://doi.org/10.3791/65326
M3 - Article
C2 - 37306462
SN - 1940-087X
VL - 2023
JO - Journal of Visualized Experiments
JF - Journal of Visualized Experiments
IS - 195
M1 - e65326
ER -