Cardiomyocyte production

1. A method for producing human cardiomyocytes comprising the steps of:

a) obtaining a selected human pluripotent stem cell clone having been prepared from a single pluripotent stem cell, the human stem cell clone being either a human embryonic stem cell clone or a human induced pluripotent stem cell clone;

b) culturing human pluripotent stem cells from the selected cell clone in a selected culture media batch, wherein the selected culture media is a maintenance culture media that maintains the human pluripotent stem cells in an undifferentiated state;

c) determining the level of one or more differentiation factors appropriate for cardiac differentiation of the selected human pluripotent stem cell clone cultured in the selected culture media batch;

d) obtaining a cardiac differentiation medium prepared by adjusting the level of the one or more differentiation factors in the cardiac differentiation medium according to the results obtained in step c); and

e) differentiating the human pluripotent stem cells cultured in step b) into cardiomyocytes by culturing the human pluripotent stem cells in the cardiac differentiation medium obtained in step d).

2. The method of claim 1 , wherein the selected human pluripotent stem cell clone is an induced pluripotent stem (iPS) cell clone.

3. The method of claim 1 , wherein the selected human pluripotent stem cell clone has been prepared from a single human pluripotent stem cell in an adherent culture.

4. The method of claim 3 , wherein the selected cell clone is obtained from a single human pluripotent stem cell by a process comprising incubating the single pluripotent stem cell in medium comprising a Rho-associated kinase (ROCK) inhibitor or a myosin II inhibitor under conditions to promote cell growth.

5. The method of claim 4 , wherein the myosin II inhibitor is blebbistatin.

6. The method of claim 1 , wherein from about 10 6 to about 10 10 of the human pluripotent stem cells are cultured in step b.

7. The method of claim 1 , wherein the human pluripotent stem cells are differentiated in a culture volume of from 5 milliliters to 25 liters.

8. The method of claim 1 , wherein the human pluripotent stem cells or cardiomyocytes differentiated therefrom or both contain one or more transgenes.

9. The method of claim 8 , wherein the one or more transgenes encode a selectable or screenable marker or both under the control of a cardiomyocyte-specific promoter.

10. The method of claim 1 , further comprising enriching or purifying the differentiated cardiomyocytes.

11. The method of claim 1 , wherein the cardiac differentiation medium comprises externally added fibroblast growth factor (FGF) in an amount of from 5 to 200 ng/ml.

12. The method of claim 1 , wherein the human pluripotent stem cells cultured in step b) are rotated or shaken at a speed of about 15 rpm to 100 rpm.

13. The method of claim 1 , wherein the culture media is TeSR, mTeSR, RPMI medium, supplemented DMEM-F12 or dilutions thereof.

14. The method of claim 1 , wherein the differentiation factors whose level are adjusted in the cardiac differentiation medium are one or more differentiation factors selected from the group consisting of a modulator of signaling pathways of bone morphogenetic protein; ActivinA/Nodal; vascular endothelial growth factor (VEGF); dickkopf homolog 1 (DKK1); basic fibroblast growth factor (bFGF); insulin growth factor (IGF); and epidermal growth factor (EGF).

15. The method of claim 14 , wherein the differentiation factors whose levels are adjusted are bone morphogenetic protein (BMP) 2, BMP4, BMP10, Activin A, bFGF, IGF, EGF, BMP signaling inhibitor, Activin signaling inhibitor, or a combination thereof.

16. The method of claim 15 , wherein the BMP signaling inhibitor is dorsomorphin.

17. The method of claim 15 , wherein the Activin A signaling inhibitor is SB431542.