Maintenance of genomic stability in cultured stem cells
The present application relates to methods and compositions for the generation of therapeutic cells having reduced incidence of karyotypic abnormalities. In several embodiments cardiac stem cells are cultured in an antioxidant-supplemented media that reduces levels of reactive oxygen species, but does not down regulate DNA repair mechanisms. In several embodiments, physiological oxygen concentrations are used during culture in order to increase the proliferation of stem cells, decrease the senescence of the cells, decrease genomic instability, and/or augment the functionality of such cells for cellular therapies.
1. A method for reducing the incidence of karyotypic abnormalities in cardiac stem cells for use in the repair or regeneration of cardiac tissue, the method comprising:
culturing cardiac stem cells in a culture media supplemented with an antioxidant composition,
wherein said antioxidant composition comprises glutathione present in a concentration ranging from 5 to 20 μM, vitamin C present in a concentration ranging from 5 to 20 μM, and vitamin E present in a concentration ranging from 5 to 20 μM.
2. The method of claim 1 , wherein said cardiac stem cells are cardiosphere-derived cells (CDCs).
3. The method according to claim 1 , wherein said glutathione is present in a concentration of 10 μM.
4. The method of claim 1 , wherein said antioxidant further comprises one or more of thiols and polyphenols.
5. The method of claim 1 , wherein said antioxidant further comprises vitamin A.
6. The method of claim 1 , wherein at least one of said vitamin C and said vitamin E is present in a concentration of 10 μM.
7. The method of claim 1 , wherein said culturing is for at least about 24 hours.
8. The method of claim 1 , wherein said at least one peptide antioxidant is glutathione, wherein said glutathione is present in a concentration ranging from 10 μM, wherein said at least one non-peptide antioxidant comprises vitamin C and vitamin E, and wherein each of said vitamin C and vitamin E is present in a concentration ranging from about 10 μM.