Differentiated hiPSC-CMs grown in a monolayer were loaded with Fura-2 AM (Thermo Fisher, Waltham, MA) at 1 μM in phenol red-free RPMI + B27 for 10 minutes at room temperature, then washed twice and allowed to de-esterify for 10 minutes at 37°C. Isoproterenol was used at a concentration of 10 nM, as previously described [23 (link)–25 (link)]. Experiments were carried out at 37°C using the IonOptix myocyte calcium and contractility system (IonOptix, Westwood, MA). Ratiometric calcium transient curves were fitted and analyzed using a custom MATLAB script.
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Measuring Calcium Dynamics in hiPSC-CMs
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Corresponding Organization : University of Minnesota Medical Center
Other organizations : Gladstone Institutes
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Variable analysis
independent variables
- Isoproterenol concentration (10 nM)
- Calcium transient curves
- Fura-2 AM concentration (1 μM)
- Loading time (10 minutes)
- De-esterification time (10 minutes)
- Temperature (37°C)
- Positive control: Not explicitly mentioned.
- Negative control: Not explicitly mentioned.
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