Method for producing mixed cell population of cardiomyocytes and vascular cells from induced pluripotent stem cell
The present invention provides: a method for producing mixed cells comprising cardiomyocytes, endothelial cells and mural cells from induced pluripotent stem cells, the method comprising (a) a step of producing cardiomyocytes from induced pluripotent stem cells and (b) a step of culturing the cardiomyocytes in the presence of VEGF; and a therapeutic agent for heart diseases, comprising the mixed cells produced by the method.
1. A myocardial sheet comprising cells derived from human induced pluripotent stem (iPS) cells, wherein a composition ratio of said cells derived from human iPS cells is 30 to 70% for cardiomyocytes, 0.1 to 20% for endothelial cells, and 1 to 40% for mural cells.
2. The myocardial sheet according to claim 1 , wherein a composition ratio of TRA-1-60-positive undifferentiated cells in the cells derived from human iPS cells is 10% or less.
3. The myocardial sheet according to claim 1 , wherein a composition ratio of TRA-1-60-positive undifferentiated cells in the cells derived from human iPS cells is 2.2% or less.
4. The myocardial sheet according to claim 1 , wherein a composition ratio of TRA-1-60-positive undifferentiated cells in the cells derived from human iPS cells is 0.1% or more.
5. A layered myocardial sheet which comprises a plurality of myocardial sheets according to claim 1 .
6. A method of treating heart disease comprising implanting the myocardial sheet according to claim 1 on the heart disease affected area of a patient suffering from heart disease.
7. A method of treating heart disease comprising implanting the layered myocardial sheet according to claim 5 on the heart disease affected area of a patient suffering from heart disease.
8. A mixed cell composition derived from human iPS cells which includes 40 to 80% of cardiomyocytes, 1 to 20% of endothelial cells and 1 to 40% of mural cells.
9. The mixed cell composition according to claim 8 , wherein a composition ratio of TRA-1-60-positive undifferentiated cells is 10% or less.
10. The mixed cell composition according to claim 8 , wherein a composition ratio of TRA-1-60-positive undifferentiated cells is 2.7% or less.
11. The mixed cell composition according to claim 8 , wherein a composition ratio of TRA-1-60-positive undifferentiated cells is 0.1% or more.
12. A method for producing the myocardial sheet according to claim 1 comprising the steps of:
culturing mixed cells including 40 to 80% of cardiomyocytes, 1 to 20% of endothelial cells and 1 to 40% of mural cells which are derived from human induced pluripotent stem cells using a culture tool coated with a temperature-responsive polymer, and
recovering the myocardial sheet.