Abstract
Human-induced pluripotent stem cells (h-iPSCs) are a unique in vitro model for cardiovascular research. To realize the potential applications of h-iPSCs-derived cardiomyocytes (CMs) for drug testing or regenerative medicine and disease modeling, characterization of the metabolic features is critical. Here, we show the transcriptional profile during stages of cardiomyogenesis of h-iPSCs-derived CMs. CM differentiation was not only characterized by the expression of mature structural components (MLC2v, MYH7) but also accompanied by a significant increase in mature metabolic gene expression and activity. Our data revealed a distinct substrate switch from glucose to fatty acids utilization for ATP production. Basal respiration and respiratory capacity in 9 days h-iPSCs-derived CMs were glycolysis-dependent with a shift towards a more oxidative metabolic phenotype at 14 and 28 day old CMs. Furthermore, mitochondrial analysis characterized the early and mature forms of mitochondria during cardiomyogenesis. These results suggest that changes in cellular metabolic phenotype are accompanied by increased O2 consumption and ATP synthesis to fulfill the metabolic needs of mature CMs activity. To further determine functionality, the physiological response of h-iPSCs-derived CMs to β-adrenergic stimulation was tested. These data provide a unique in vitro human heart model for the understanding of CM physiology and metabolic function which may provide useful insight into metabolic diseases as well as novel therapeutic options.
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Abbreviations
- β-AR:
-
β-Adrenergic receptor
- CD36:
-
Cluster of differentiation 36
- CM:
-
Cardiomyocyte
- CPT-1B:
-
Carnitine-palmitoyl-CoA transferase-1B
- DGAT1:
-
Diglyceride acyltransferase 1
- DRP1:
-
Dynamin-related protein 1
- ESC:
-
Embryonic stem cells
- FA:
-
Fatty acid
- FAO:
-
Fatty acid oxidation
- FCM:
-
Flow cytometry
- GLUT:
-
Glucose transporters
- h-ESCs:
-
Human-embryonic stem cells
- h-iPSCs:
-
Human-induced pluripotent stem cells
- MFF:
-
Mitochondrial fission factor
- MFN2:
-
Mitofusin-2
- MLC2a:
-
Myosin light chain 2 isoform atrial
- MLC2v:
-
Myosin light chain 2 isoform ventricular
- OPA1:
-
Optic atrophy 1
- PDK4:
-
Pyruvate dehydrogenase kinase 4
- PFA:
-
Paraformaldehyde
- PGC-1α:
-
PPARγ coactivator-1α
- PP2A:
-
Protein phosphatase 2A
- PPARα:
-
Peroxisome proliferator-activated receptor-α
- ROS Reactive:
-
Reactive oxygen species
- SM:
-
Standard manufacturer’s maintenance medium
- TEM:
-
Transmission electron microscopy
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Acknowledgements
We thank Dr. Martin Förster, for his support during FCM procedures and Beate Schulze for her technical assistance, Department of Cardiology, University Hospital Jena.
Funding
This study was supported by the Interdisciplinary Center for Clinical Research (IZKF, J55) of the Medical Faculty, University of Jena.
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Bekhite, M.M., González Delgado, A., Menz, F. et al. Longitudinal metabolic profiling of cardiomyocytes derived from human-induced pluripotent stem cells. Basic Res Cardiol 115, 37 (2020). https://doi.org/10.1007/s00395-020-0796-0
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DOI: https://doi.org/10.1007/s00395-020-0796-0