miR-27b and miR-23b Modulate Cardiomyocyte Differentiation from Mouse Embryonic Stem Cells
Abstract
:1. Introduction
2. Material and Methods
2.1. ESC Culture and microRNA Transfection Assays
2.2. RNA Isolation, Reverse Transcriptase Reaction and Quantitative Real Time PCR
Primers | Sequence | Annealing Temperature | Amplicon size |
---|---|---|---|
Wnt3a | Sense: 5'-TTCTGCAGGAACTACGTGGA-3' | 60 °C | 179 bp |
Brachyury | Sense: 5'- CGACCACAAAGATGTAATGGAG-3' | 59.5 °C | 120 bp |
Kcnk3 | Sense: 5'-TGTGCACCTTCACCTACCTG-3' | 59.7 °C | 203 bp |
Kcnh1 | Sense: 5'-ACGCCCTTCAGAAAGTGCTA-3' | 62 °C | 182 bp |
c-Troponin T | Sense: 5'-TTCGACCTGCAGGAAAAGTT-3' | 64 °C | 133bp |
Hcn4 | Sense: 5'-CAGCGTCAGAGCGGATACTT-3' | 60 °C | 158bp |
Cx40 | Sense: 5'-CAGAGCCTGAAGAAGCCAAC-3' | 60 °C | 178bp |
Cx43 | Sense: 5'-GAGAGCCCGAACTCTCCTTT-3' | 60 °C | 158bp |
miR 27b | 5'-UUCACAGUGGCUAAGUUCUGC-3' | 60 °C | 21 bp |
miR 23b | 5'-AUCACAUUGCCAGGGAUUACC-3' | 60 °C | 21 bp |
3. Results
3.1. miR-27b Inhibits in Vitro Cardiac Differentiation by Modulating Wnt-Beta Catenin Canonical Pathway
3.2. miR-23b Regulates the Beating Phenotype During in Vitro Cardiac Differentiation from ESCs
4. Discussion
5. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Vilches, J.M.; Pulido, A.; Hernández-Torres, F.; Franco, D.; Aránega, A. miR-27b and miR-23b Modulate Cardiomyocyte Differentiation from Mouse Embryonic Stem Cells. J. Cardiovasc. Dev. Dis. 2014, 1, 41-51. https://doi.org/10.3390/jcdd1010041
Vilches JM, Pulido A, Hernández-Torres F, Franco D, Aránega A. miR-27b and miR-23b Modulate Cardiomyocyte Differentiation from Mouse Embryonic Stem Cells. Journal of Cardiovascular Development and Disease. 2014; 1(1):41-51. https://doi.org/10.3390/jcdd1010041
Chicago/Turabian StyleVilches, José Manuel, Antonio Pulido, Francisco Hernández-Torres, Diego Franco, and Amelia Aránega. 2014. "miR-27b and miR-23b Modulate Cardiomyocyte Differentiation from Mouse Embryonic Stem Cells" Journal of Cardiovascular Development and Disease 1, no. 1: 41-51. https://doi.org/10.3390/jcdd1010041