Related Experiment Video
Updated: Apr 12, 2026
Generation of Ventricular-Like HiPSC-Derived Cardiomyocytes and High-Quality Cell Preparations for Calcium Handling Characterization
Published on: January 17, 2020
Comparable calcium handling of human iPSC-derived cardiomyocytes generated by multiple laboratories
Hyun Seok Hwang1, Dmytro O Kryshtal1, T K Feaster2
1Division of Clinical Pharmacology, Oates Institute for Experimental Therapeutics, Vanderbilt University School of Medicine, Nashville, TN, USA.
Insights
Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) show functional calcium handling comparable to adult cells. These hiPSC-CMs, generated via monolayer methods, are reproducible and suitable for disease modeling.
Area of Science:
- Cardiology
- Stem Cell Biology
- Biophysics
Background:
- Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) are valuable for modeling heart diseases.
- Traditional aggregation methods yield hiPSC-CMs with immature sarcoplasmic reticulum (SR) Ca stores.
- Novel monolayer differentiation methods produce high-purity hiPSC-CMs, but their calcium handling requires thorough investigation.
Purpose of the Study:
- To comprehensively investigate the calcium (Ca) handling and cytosolic Ca buffering properties of hiPSC-CMs generated using monolayer-based methods.
- To compare the Ca handling of hiPSC-CMs from multiple independent laboratories and hiPSC lines with adult cardiomyocytes.
- To assess the reproducibility of hiPSC-CMs after cryopreservation and recovery.
Main Methods:
- hiPSC-CMs were generated independently at three universities using monolayer-based differentiation protocols.
- Cells were cryopreserved, shipped, recovered, and plated for functional analysis.
- Calcium handling properties, including L-type Ca currents, Ca transients, SR Ca stores, SERCA, and NCX activity, were compared to adult rabbit and mouse ventricular cardiomyocytes.
Main Results:
- hiPSC-CMs exhibited robust L-type Ca currents, twitch Ca transients, and caffeine-releasable SR Ca stores comparable to adult CMs.
- Cytoplasmic Ca buffering and cell capacitance to cell volume ratio were similar to adult rabbit CMs, despite smaller cell volume.
- Sarcoendoplasmic reticulum Ca ATPase (SERCA) and Na/Ca exchanger (NCX) transport were functional and comparable in relative contribution to adult CMs, though slower.
- Ca handling maturity increased with differentiation time (15-21 days post-induction).
Conclusions:
- hiPSC-CMs generated via monolayer methods from multiple iPSC lines demonstrate reproducible cryopreservation and recovery.
- These hiPSC-CMs possess functional sarcoplasmic reticulum (SR) Ca handling capabilities comparable to adult cardiomyocytes.
- Monolayer-derived hiPSC-CMs are a reliable and functional model for studying human cardiac physiology and disease.
Abstract:
Cardiomyocytes (CMs) derived from human induced pluripotent stem cells (hiPSCs) are being increasingly used to model human heart diseases. hiPSC-CMs generated by earlier aggregation-based methods (i.e., embryoid body) often lack functional sarcoplasmic reticulum (SR) Ca stores characteristic of mature mammalian CMs. Newer monolayer-based cardiac differentiation methods (i.e., Matrigel sandwich or small molecule-based differentiation) produce hiPSC-CMs of high purity and yield, but their Ca handling has not been comprehensively investigated. Here, we studied Ca handling and cytosolic Ca buffering properties of hiPSC-CMs generated independently from multiple hiPSC lines at Stanford University, Vanderbilt University and University of Wisconsin-Madison. hiPSC-CMs were cryopreserved at each university. Frozen aliquots were shipped, recovered from cryopreservation, plated at low density and compared 3-5days after plating with acutely-isolated adult rabbit and mouse ventricular CMs. Although hiPSC-CM cell volume was significantly smaller, cell capacitance to cell volume ratio and cytoplasmic Ca buffering were not different from rabbit-CMs. hiPSC-CMs from all three laboratories exhibited robust L-type Ca currents, twitch Ca transients and caffeine-releasable SR Ca stores comparable to adult CMs. Ca transport by sarcoendoplasmic reticulum Ca ATPase (SERCA) and Na/Ca exchanger (NCX) was similar in all hiPSC-CM lines, but slower compared to rabbit-CMs. However, the relative contribution of SERCA and NCX to Ca transport of hiPSC-CMs was comparable to rabbit-CMs. Ca handling maturity of hiPSC-CMs increased from 15 to 21days post-induction. We conclude that hiPSC-CMs generated independently from multiple iPSC lines using monolayer-based methods can be reproducibly recovered from cryopreservation and exhibit comparable and functional SR Ca handling.
More Related Videos
06:42Author Spotlight: Real-Time Measurements of Calcium and Contractility Parameters in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes
Published on: May 26, 2023
10:01High-Throughput Optical Controlling and Recording Calcium Signal in iPSC-Derived Cardiomyocytes for Toxicity Testing and Phenotypic Drug Screening
Published on: March 31, 2022