to CMs [8, 9]. This protocol aims to evaluate cell lines for their

cardiac differentiation potential as well as describe an integrated

system for their production using MC cultures.

2

Materials

Here lists the minimal equipment, medium, chemicals, and

reagents required for culture, differentiation, and characterization

of hiPSCs toward CMs.

2.1

General

Equipment and

Consumables

1. Biosafety cabinet Class II, Type A2 (BSC).

2. CO2 incubator.

3. Eppendorf^® New Brunswick™S41i CO2 Incubator Shaker.

4. Corning^® Four Position Magnetic Stirrer.

5. Pipettors.

6. Aspiration device.

7. Bench rocker.

8. Low-speed centrifuge (for spinning cells).

9. Laboratory water bath.

10. Phase contrast microscope.

11. NucleoCounter^® automated cell counter (Chemometec).

12. Flow cytometer.

13. 15-mL and 50-mL conical tubes.

14. 40-μm Cell Strainer.

15. Tissue culture-treated six-well plates.

16. Tissue culture-treated 24-well plates.

17. 0.2 μm PES filter (for media filtration).

2.2

Media and

Consumables for

hiPSC Cultures

1. 60 mm tissue culture-treated culture dish.

2. Tissue culture-treated six-well plates.

3. mTeSR™1 (StemCell Technologies).

4. DMEM/F12 with glutamine and HEPES (Gibco).

5. GelTrex™(Gibco).

6. Dulbecco’s phosphate-buffered saline without Ca²⁺ and Mg²⁺,

DPBS(
) (Gibco).

7. TrypeLE-Express (Gibco).

8. ReLeSR™(StemCell Technologies).

9. Y27632 (Rock inhibitor, RI) (StemCell Technologies).

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