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Updated: Aug 25, 2025
Modified Mouse Embryonic Stem Cell based Assay for Quantifying Cardiogenic Induction Efficiency
Published on: April 22, 2011
Inducible apelin receptor knockdown reduces differentiation efficiency and contractility of hESC-derived
Robyn G C Macrae1,2, Maria T Colzani2, Thomas L Williams1
1Experimental Medicine and Immunotherapeutics, University of Cambridge, Addenbrooke's Hospital, Level 6, Addenbrooke's Centre for Clinical Investigation, Box 110, Cambridge CB2 0QQ, UK.
Insights
Researchers developed a new model using human stem cell-derived cardiomyocytes to study the apelin receptor
Area of Science:
- Cardiovascular Biology
- Stem Cell Research
- G protein-coupled receptor signaling
Background:
- The apelin receptor is crucial for cardiovascular function and disease.
- Existing human in vitro models are insufficient for studying the apelinergic system in heart cells.
Purpose of the Study:
- To establish a human in vitro model using stem cell-derived cardiomyocytes to investigate the apelin receptor's role.
- To examine the impact of apelin receptor function loss on cardiomyocyte development and disease.
Main Methods:
- Generated human embryonic stem cell-derived cardiomyocytes (hESC-CMs) with an inducible apelin receptor knockdown system.
- Assessed apelin receptor and ligand expression in hESCs and hESC-CMs.
- Utilized phenotypic assays on differentiated hESC-CMs and 3D engineered heart tissues.
Main Results:
- Apelin receptor knockdown during differentiation impaired cardiomyocyte development and led to asynchronous contraction.
- Knockdown in engineered heart tissues reduced contractility and increased stiffness, mimicking disease phenotypes.
- hESC-CMs expressed the apelin signaling system comparably to adult heart tissue.
Conclusions:
- Successfully demonstrated the apelin receptor's critical role in hESC-CM differentiation using an inducible knockdown system.
- Developed a 3D engineered heart tissue model that recapitulates apelin receptor down-regulation in heart failure.
- This model offers a platform for studying heart failure and evaluating new therapies.
Aims:
The apelin receptor, a G protein-coupled receptor, has emerged as a key regulator of cardiovascular development, physiology, and disease. However, there is a lack of suitable human in vitro models to investigate the apelinergic system in cardiovascular cell types. For the first time we have used human embryonic stem cell-derived cardiomyocytes (hESC-CMs) and a novel inducible knockdown system to examine the role of the apelin receptor in both cardiomyocyte development and to determine the consequences of loss of apelin receptor function as a model of disease.
Methods And Results:
Expression of the apelin receptor and its ligands in hESCs and hESC-CMs was determined. hESCs carrying a tetracycline-inducible short hairpin RNA targeting the apelin receptor were generated using the sOPTiKD system. Phenotypic assays characterized the consequences of either apelin receptor knockdown before hESC-CM differentiation (early knockdown) or in 3D engineered heart tissues as a disease model (late knockdown). hESC-CMs expressed the apelin signalling system at a similar level to the adult heart. Early apelin receptor knockdown decreased cardiomyocyte differentiation efficiency and prolonged voltage sensing, associated with asynchronous contraction. Late apelin receptor knockdown had detrimental consequences on 3D engineered heart tissue contractile properties, decreasing contractility and increasing stiffness.
Conclusions:
We have successfully knocked down the apelin receptor, using an inducible system, to demonstrate a key role in hESC-CM differentiation. Knockdown in 3D engineered heart tissues recapitulated the phenotype of apelin receptor down-regulation in a failing heart, providing a potential platform for modelling heart failure and testing novel therapeutic strategies.
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