Methods and compositions for maintaining genomic stability in cultured stem cells

摘要

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.

主权项

1. A method for culturing cardiac stem cells for use in the repair or regeneration of cardiac tissue while reducing DNA damage during culturing, the method comprising:
isolating cardiac stem cells, wherein said cardiac stem cells are isolated from healthy mammalian non-embryonic cardiac tissue; and culturing said isolated cardiac stem cells as a monolayer culture in a culture media comprising an antioxidant composition, said antioxidant composition comprising glutathione present in a concentration ranging from 5 to 20 μM, vitamin C present in a concentration ranging from 0.1 to 20 μM, and vitamin E present in a concentration ranging from 0.1 to 20 μM, wherein said antioxidant composition present in said culture media is sufficient to (i) reduce reactive oxygen species to a level which decreases oxidative stress induced DNA damage, (ii) not reduce markers of DNA repair mechanisms to a statistically significant degree, and (iii) not induce markers of DNA damage in said cardiac stem cells to a statistically significant degree, when compared to cardiac stem cells not contacted with said antioxidant composition.