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High Efficiency Differentiation of Human Pluripotent Stem Cells to Cardiomyocytes and Characterization by Flow Cytometry
Published on: September 23, 2014
Calcium handling in human embryonic stem cell-derived cardiomyocytes
Jonathan Satin1, Ilanit Itzhaki, Sophia Rapoport
1The Sohnis Laboratory for Cardiac Electrophysiology and Regenerative Medicine, Rappaport Faculty of Medicine, Technion-Israel Institute of Technology, Haifa, Israel.
Developing human embryonic stem cell-derived cardiomyocytes (hESC-CMs) exhibit functional sarcoplasmic reticulum calcium stores and unique calcium handling patterns. This research is vital for understanding cardiomyocyte development and future cell replacement therapies.
Area of Science:
- Cardiology
- Stem Cell Biology
- Cell Physiology
Background:
- Human embryonic stem cell-derived cardiomyocytes (hESC-CMs) are crucial for regenerative medicine.
- Understanding calcium handling is fundamental to cardiomyocyte function and development.
Purpose of the Study:
- To characterize calcium handling mechanisms in developing hESC-CMs.
- To investigate the role of sarcoplasmic reticulum (SR) and inositol-1,4,5-trisphosphate (IP3) receptors in hESC-CM calcium regulation.
Main Methods:
- Real-time PCR, immunocytochemistry, and whole-cell voltage-clamp electrophysiology.
- Simultaneous patch-clamp/confocal calcium imaging and surface membrane labeling.
- Caffeine application and caged-IP3 photolysis to assess calcium store function.
Main Results:
- hESC-CMs possess functional SR calcium stores with ryanodine receptor-2 and IP3 receptors.
- Action-potential-induced calcium transients propagate as waves, with localized surface membrane sparks observed.
- Calcium store load increases with in vitro maturation, and IP3-sensitive calcium pools are functional.
Conclusions:
- Early-stage hESC-CMs display a functional SR calcium store and distinct calcium handling.
- Functional characterization of hESC-CMs is essential for developmental research and myocardial regeneration strategies.
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