Table 2.
Main aspects of cardiomyocyte differentiation methods from ESCs and iPSCs.
| Reference | Guan53 | Lin55 | Hashimoto56 |
|---|---|---|---|
| Starting cells | iPSCs from urine cells | iPSCs from dermal fibroblasts | iPSCs from T lymphocytes |
| Proliferation media | mTeSR | DMEM / F12 | DMEM / F12 |
| Factors | not used | KSR, bFGF | KSR |
| Duration (days) | 4 | until 30–40 passages | 20 |
| Cardiomyocytes differentiation | |||
| EB acquisition | not used | by dissociation | by dissociation |
| Differentiation medium | RPMI | basal StemPro-34 | basal StemPro-34 |
| Factors | Activin A, BMP4, 2% B27 minus insulin, L-glutamine |
Activin A, BMP4, bFGF, XAV-939, VEGF-A |
KSR, transferrin, CHIR99021, IDE1, bFGF, IWP-2, VEGF, SB431542, dorsomorphin |
| Duration (days) | 12 | 22 | 20 |
| Purification | Not used | Not used | Metabolic by glucose-free DMEM plus L-lactate (8 days) |
| Protein expression | Cardiac α and β MHC, connexin 43, DMD | DMD, CTNT |
Sarcomeric α-actinin, MLC2v |
| Differentiation markers | Nkx2.5, ACTN1 | CTNT | ACTN1, MLC2v |
| Advantages | Rapid differentiation protocol; no FACS; serum-free protocol |
Efficient differentiation by small molecules; no FACS | Differentiation by small molecules; purification for high quality cardiomyocytes |
| Disadvantages | Ethical problems for exogenous growth factor usage | Ethical problems for exogenous factors; longer time range to reach adequate number of cardiomyocytes; serum medium |
Ethical problems for exogenous factors; lower amount of cardiomyocytes; serum medium |
ACTN1, alpha-actinin-1; basal StemPro-34, basal serum-free medium plus nutrient supplement; bFGF, basic fibroblast growth factor; BMP4, bone morphogenetic protein 4; CTNT, cardiac troponin T; DMD, Duchenne muscular dystrophy; DMEM, Dulbecco’s modified Eagle medium; ESC, embryonic stem cell; FACS, fluorescence-activated cell sorting; IDE1, inducer of definitive endoderm 1; iPSC, induced pluripotent stem cell; IWP, inhibitor of Wnt processing; KSR, knockout serum replacement; MHC, myosin heavy chain; MLC2v, myosin light chain 2v; mTeSR, cGMP feeder-free maintenance medium for human ESCs and iPSCs; Nkx2.5, Homeobox NKX2.5; RPMI, Roswell Park Memorial Institute; VEGF-A, vascular endothelial growth factor A.