Methods for inducing cell death in pluripotent stem cells and differentiated cells other than cardiac myocytes
View Patent ↗The present invention has as its object developing a method that does not involve genomic modification and which yet is capable of inducing cell death in pluripotent stem cells such as embryonic stem cells and induced pluripotent stem cells, as well as in differentiated cells other than cardiomyocytes derived from pluripotent stem cells, but not in cardiomyocytes. It has been revealed that by establishing a method capable of inducing cell death in cells other than cardiomyocytes in a very efficient manner by adding a substance having no recognized inherent toxicity or cell death inducing action to the culture conditions for pluripotent stem cells and non-cardiomyocytes, the stated problem can be solved without relying upon genomic modification.
1. A method for selecting for cardiomyocytes derived from pluripotent stem cells comprising culturing a population of cells including pluripotent stem cells, cardiomyocytes, and other cells derived from the pluripotent stem cells in a hypertonic solution having an osmotic pressure of 370 mOsm/kg or higher, wherein cell death is induced in non-cardiomyocyte cells.
2. The method according to claim 1 , wherein culturing is conducted in the hypertonic solution for 2 hours or longer.
3. The method according to claim 1 , wherein the hypertonic solution is prepared by adding a saccharide to a culture medium.
4. The method according to claim 3 , wherein the hypertonic solution comprises 0.1-1 M saccharides.
5. The method according to claim 3 , wherein saccharides are sugar alcohols, sugars, or betaines.
6. The method according to claim 5 , wherein the sugar alcohols, sugars, or betaines are mannitol, sorbitol, xylitol, glycerol, sucrose, glucose, trimethylglycine, or combinations thereof.
7. The method according to claim 4 , wherein the hypertonic solution comprises 0.1-0.6 M glycerol.
8. The method according to claim 7 , wherein culturing is conducted for 10 hours or longer.
9. The method according to claim 1 , further comprising returning the cell population, that has been cultured in the hypertonic solution, to a culturing medium having the normal osmotic pressure of 200-300 mOsm/kg and subjecting the cell population to further culturing.
10. The method of claim 3 , wherein the saccharide is a carbohydrate.
11. A method for inducing cell death in a cell population comprising pluripotent stem cells and cardiomyocytes derived from pluripotent stem cells comprising culturing said cell population in a hypertonic solution having an osmotic pressure of 370 mOsm/kg or higher,
wherein said pluripotent stem cells die and the cardiomyocytes derived from pluripotent stem cells do not die.