Methods for reprogramming differentiated non-cardiac cells into cardiomyocytes
Compositions and methods are described herein for chemically inducing cells to change their differentiation state and become cardiac progenitor cells or cardiomyocytes.
1. A method of generating a cardiac progenitor cell or a cardiomyocyte comprising:
contacting at least one fibroblast or a cell derived therefrom with a cardiac induction medium comprising a GSK3 inhibitor, a BMP4 agonist, an Activin A agonist and a VEGF agonist; and
allowing the cells to incubate in the medium;
thereby generate at least one cardiac progenitor cell or a cardiomyocyte.
2. The method of claim 1 , wherein the fibroblast is a human fibroblast.
3. The method of claim 1 , wherein the fibroblast is contacted with one or more chemical agents prior to the contacting with the cardiac induction medium.
4. The method of claim 2 , wherein the fibroblast is contacted with one or more agents selected from a WNT agonist, a GSK3 inhibitor, a TGF-beta inhibitor, an inhibitor of extracellular signal-regulated kinase 1 (ERK1), an inhibitor of Ras GTPase-activating protein (Ras-GAP), an Oct-4 activator, a rho-associated protein kinase, an iron chelator, a KDMSB inhibitor, a histone methyltransferase inhibitor, a PDGF tyrosine kinase inhibitor.
5. The method of claim 4 , wherein the fibroblast is contacted with each agent for a time sufficient to induce the cardiac progenitor cell or cardiomyocyte to express α-Actinin, MLC2v, MY20, cMHC NKX2-5, MEF2c, GATA4, ISLL cTNL cTN1, MLC2a or any combination thereof.
6. The method of claim 1 , furthering comprising administering the cardiac progenitor cell(s) or the cardiomyocyte(s) to a subject.
7. The method of claim 1 , wherein the fibroblast is allogenic.
8. The method of claim 1 , wherein the fibroblast is autologous.
9. The method of claim 1 , wherein the fibroblast is obtained from a subject who suffers or is suspected of suffering from a heart condition or disease.
10. The method of claim 1 , wherein the fibroblast is obtained from a healthy subject.
11. The method of claim 1 , wherein the cardiac progenitor cell or the cardiomyocyte is allogenic.
12. The method of claim 1 , wherein the cardiac progenitor cell or the cardiomyocyte is autologous.
13. A method of generating a cardiac progenitor cell or a cardiomyocyte comprising:
contacting at least one fibroblast with a cardiac reprogramming medium comprising one or more agents selected from a WNT agonist, a GSK3 inhibitor, a TGF-beta inhibitor, an inhibitor of extracellular signal-regulated kinase 1 (ERK1), an inhibitor of Ras GTPase-activating protein (Ras-GAP), an Oct-4 activator, a rho-associated protein kinase, an iron chelator, a KDMSB inhibitor, a histone methyltransferase inhibitor, a PDGF tyrosine kinase inhibitor;
contacting the cell with a cardiac induction medium comprising a GSK3 inhibitor, a BMP4 agonist, an Activin A agonist and a VEGF agonist;
thereby generating at least one cardiac progenitor cell or a cardiomyocyte.
14. The method of claim 13 , wherein the fibroblast is a human fibroblast.
15. The method of claim 13 , wherein the fibroblast is contacted with each agent for a time sufficient to induce the cardiac progenitor cell or cardiomyocyte to express α-Actinin, MLC2v, MY20, cMHC NKX2-5, MEF2c, GATA4, ISLL cTNL cTN1, MLC2a or any combination thereof.
16. The method of claim 13 , furthering comprising administering the cardiac progenitor cell or the cardiomyocyte to a subject.
17. The method of claim 10 , wherein the fibroblast is allogenic.
18. The method of claim 10 , wherein the fibroblast is autologous.
19. The method of claim 10 , wherein the fibroblast is obtained from a subject who suffers or is suspected of suffering from a heart condition or disease.
20. The method of claim 10 , wherein the fibroblast is obtained from a healthy subject.
21. The method of claim 10 , wherein the cardiac progenitor cell or the cardiomyocyte is allogenic.
22. The method of claim 10 wherein the cardiac progenitor cell or the cardiomyocyte is autologous.