Histone Deacetylase Inhibitor Suberoylanilide Hydroxamic Acid Improves Energetic Status and Cardiomyogenic Differentiation of Human Dilated Myocardium-Derived Primary Mesenchymal Cells
Abstract
:1. Introduction
2. Results
2.1. Isolation and Identification of Healthy and Pathological hmMSCs
2.2. Energetic Profile of Healthy and Pathological hmMSCs
2.3. Histone Deacetylase Activity and Effect of SAHA on Healthy and Pathological hmMSC
2.4. The Impact of SAHA on Bioenergetics of Healthy and Pathological Cells
2.5. The Impact of SAHA on Cardiomyogenic Differentiation
3. Discussion
4. Materials and Methods
4.1. Isolation and Growth of hmMSC
4.2. Flow Cytometry
4.3. Measurement of Cell Proliferation
4.4. HDAC Activity Assay
4.5. ATP Activity Assay
4.6. Measurement of Mitochondrial Membrane Potential
4.7. Real Time PCR (RT-PCR)
4.8. Measurement of Metabolic Activity with Seahorse
4.9. Western Blotting
4.10. Immunocytochemistry
4.11. Electron Microscopy
4.12. Statistics
4.13. Ethical Approval
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
DCM | Dilated cardiomyopathy |
HDAC | Histone deacetylase |
JC1 | 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimi- dazolylcarbocyanine iodide |
HDACi | Histone deacetylase inhibitor |
SAHA | Suberoylanilide hydroxamic acid |
ATP | Adenosine triphosphate |
iPS | Induced pluripotent stem cells |
RNA | Ribonucleic acid |
ECAR | Extracellular acidification |
OCR | Oxygen consumption rate |
FCCP | Carbonyl cyanide-4-(trifluoromethoxy)phenylhydrazone |
2-DG | 2-Deoxy-D-glucose |
ACTC1 | Actin alpha cardiac muscle 1 |
ACTB | Actin beta |
GAPDH | Glyceraldehyde 3-phosphate dehydrogenase |
DAPI | 4′,6-diamidino-2-phenylindole |
ROS | Reactive oxygen species |
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Miksiunas, R.; Rucinskas, K.; Janusauskas, V.; Labeit, S.; Bironaite, D. Histone Deacetylase Inhibitor Suberoylanilide Hydroxamic Acid Improves Energetic Status and Cardiomyogenic Differentiation of Human Dilated Myocardium-Derived Primary Mesenchymal Cells. Int. J. Mol. Sci. 2020, 21, 4845. https://doi.org/10.3390/ijms21144845
Miksiunas R, Rucinskas K, Janusauskas V, Labeit S, Bironaite D. Histone Deacetylase Inhibitor Suberoylanilide Hydroxamic Acid Improves Energetic Status and Cardiomyogenic Differentiation of Human Dilated Myocardium-Derived Primary Mesenchymal Cells. International Journal of Molecular Sciences. 2020; 21(14):4845. https://doi.org/10.3390/ijms21144845
Chicago/Turabian StyleMiksiunas, Rokas, Kestutis Rucinskas, Vilius Janusauskas, Siegfried Labeit, and Daiva Bironaite. 2020. "Histone Deacetylase Inhibitor Suberoylanilide Hydroxamic Acid Improves Energetic Status and Cardiomyogenic Differentiation of Human Dilated Myocardium-Derived Primary Mesenchymal Cells" International Journal of Molecular Sciences 21, no. 14: 4845. https://doi.org/10.3390/ijms21144845