Cardiomyocyte Differentiation

This is a formatted protocol for differentiating human induced pluripotent stem cells (hiPSCs) into cardiomyocytes. The protocol is based on the small-molecule method published in Burridge et al. Nat Methods 2014 and has been used extensively in publications at the Stanford Cardiovascular Institute and elsewhere.

Differentiation of hiPSCs into Cardiomyocytes

Day 0

Mesoderm Induction
CHIR-99021
10 min
1. Take iPSC cells (80% confluency). Remove E8 medium.
2. Add RPMI +B27 (-)insulin +CHIR-99021 (6 µM).
Day 2

Recovery

10 min
1. Change media to RPMI+B27 (-) insulin.
Day 3

Cardiac Progenitor
IWR-1-endo
10 min
1. Change media to RPMI +B27 (-)insulin +IWR-1-endo (5 µM).
Day 5

Recovery

10 min
1. Change media to RPMI+B27 (-) insulin.
Day 7

Early CM

10 min
1. Change media to RPMI + B27 + insulin
Day 9

Early CM

10 min
1. Change media to 3 mL RPMI + B27 +insulin.
Day 11

Early CM

10 min
1. Change media to 3 mL RPMI + B27 +insulin.
Day 12

Serum Starvation

10 min
1. Change media to 3 mL RPMI+B27 -glucose +insulin.

Cardiomyocyte Differentiation

Differentiation of hiPSCs into Cardiomyocytes

Day 0

Mesoderm Induction

Day 2

Recovery

Day 3

Cardiac Progenitor

Day 5

Recovery

Day 7

Early CM

Day 9

Early CM

Day 11

Early CM

Day 12

Serum Starvation