Simultaneous Optical Imaging of Action Potentials and Calcium Transients in Human Induced Pluripotent Stem...

Area of Science:

• Cardiology
• Stem Cell Biology
• Pharmacology

Background:

• Cardiovascular diseases are a leading cause of mortality.
• Drug discovery for cardiovascular diseases is limited by the lack of suitable in vitro platforms.
• Human induced pluripotent stem cells (hiPSCs) differentiated into cardiomyocytes (hiPSC-CMs) offer a promising model, but require functional monitoring.

Purpose of the Study:

• To develop and present a comprehensive protocol for the simultaneous optical measurement of action potentials (APs) and intracellular calcium (Ca2+) transients in hiPSC-CMs.
• To establish a high-throughput platform for cardiovascular disease modeling and drug screening.
• To facilitate the discovery of novel compounds targeting cardiovascular diseases characterized by abnormal APs and Ca2+ handling.

Main Methods:

• Adaptation of the IonOptix system for simultaneous optical detection.
• Loading hiPSC-CMs with fluorescent dyes FluoVolt (for APs) and Rhod 2 (for Ca2+ transients).
• Detailed protocols for hiPSC-CM preparation, device setup, optical imaging, and data analysis.

Main Results:

• Successful simultaneous measurement of APs and Ca2+ transients in hiPSC-CMs.
• Demonstration of a powerful high-throughput platform for functional assessment of hiPSC-CMs.
• Establishment of a comprehensive protocol for researchers.

Conclusions:

• The developed optical imaging system provides a robust platform for monitoring hiPSC-CM function.
• This method aids in the discovery of new drugs for cardiovascular diseases by analyzing cellular phenotypes.
• The protocol enables efficient and simultaneous assessment of critical cardiomyocyte parameters.