Methods of isolating non-senescent cardiac stem cells and uses thereof
The invention describes the isolation and methods of use of a non-senescent pool of adult cardiac stem cells. In particular, a subset of adult cardiac stem cells with superior regenerative capacity is disclosed. Such cells were found to have immortal DNA. Compositions comprising the non-senescent stem cells are also described. In addition, the present invention provides methods for repairing aged myocardium or damaged myocardium using the isolated non-senescent adult cardiac stem cells.
1. A pharmaceutical composition comprising a population of isolated human adult c-kit positive cardiac stem cells and a pharmaceutically acceptable carrier, wherein at least 50% of said isolated cardiac stem cells have immortal DNA.
2. The composition of claim 1 , wherein the cardiac stem cells are IGF-1 receptor positive.
3. The composition of claim 1 , wherein the cardiac stem cells have telomeres greater than 5 kbp in length.
4. The composition of claim 3 , wherein the cardiac stem cells have telomeres greater than 7 kbp in length.
5. The composition of claim 1 , wherein the cardiac stem cells are p16 INK4a negative.
6. The composition of claim 1 , wherein the cardiac stem cells are isolated from human myocardial tissue.
7. The composition of claim 1 , wherein the population is obtained by selecting cardiac stem cells that are bromodeoxyuridine (BrdU) negative following at least one passage of the cells in culture after BrdU labeling.
8. A method of repairing damaged myocardium in a subject in need thereof by administering to the subject a composition of claim 1 , wherein the structural and functional integrity of the damaged myocardium is at least partially restored following administration of the composition.
9. The method of claim 8 , wherein the isolated adult cardiac stem cells are autologous.
10. The method of claim 8 , wherein the composition is administered intramyocardially or intracoronarily.
11. The composition of claim 1 , wherein at least 80% of the isolated cardiac stem cells have immortal DNA.
12. The composition of claim 1 , wherein the cardiac stem cells are negative for markers of the cardiac, skeletal muscle, neural, and hematopoietic cell lineages.
13. An isolated population of human adult c-kit positive cardiac stem cells, wherein at least 50% of the cardiac stem cells have immortal DNA, and wherein the population is obtained by
labeling human adult c-kit positive cardiac stem cells with a DNA label, culturing the labeled cardiac stem cells for at least one population doubling in the absence of the DNA label, and selecting the cardiac stem cells that are negative for the DNA label.
14. The cell population of claim 13 , wherein the DNA label is BrdU, chlorodeoxyuridine (CldU), or iododeoxyuridine (IdU).
15. The cell population of claim 13 , wherein the cardiac stem cells are IGF-1 receptor positive and p16 INK4a negative.
16. The cell population of claim 13 , wherein the cardiac stem cells are negative for markers of the cardiac, skeletal muscle, neural, and hematopoietic cell lineages.
17. The cell population of claim 13 , wherein at least 80% of the cardiac stem cells in the population have immortal DNA.
18. The cell population of claim 13 , wherein 100% of the cardiac stem cells in the population have immortal DNA.
19. The cell population of claim 13 , wherein the cardiac stem cells are isolated from human myocardial tissue.
20. The composition of claim 1 , wherein 100% of the isolated cardiac stem cells have immortal DNA.
21. The cell population of claim 13 , wherein the cardiac stem cells in the population divide by non-random chromatid segregation when expanded in culture.