Table 2.
Factors and media used to differentiate pluripotent stem cells to cardiomyocytes.
| Different-iation method cell type | Growth factor or small molecules | Culture media | Glucose concent-ration | % of Cardiac markers | References |
|---|---|---|---|---|---|
| EB-Method hESCs iPSc |
Cells in suspension -optionally differentiated with serum-based media, DMSO, all-trans retinoic acid or 5-Aza |
80% KO-DMEM, 1 mMol L-glutamine, 1.1-1.2mMol B-ME,1% NEAA, 20% FBS |
4.5mM | 8.1% spontaneously beating EBs. In beating EBs, 29.4% cTnI+ cells. 25% spontaneously beating EBs by day 8 and 70% by day 16 of differentiation 5-Aza enhanced levels of cardiac α-MHC 10 to 15% positive for sarcomeric MHC |
(204) (215) (216) (217) (218) |
| Monolayer hESCs hiPSC |
Guided differentiation: h-Activin A, h-BMP4 +/- staged addition of h-Wnt3a & h-DKK1 |
RPMI+B27 | 11.1mM Note: 2uM of oleic acid |
> 30% CMs Differentiated cells underwent Percoll gradient centrifuge for CM enrichment (69 ± 10% CMs) Increase in sarcomeric MHC+ cells from 4 to 27% |
(27) (70) (208) |
| EB-Method hESCs hiPSC |
END-2 method (Insulin depletion, PGI2, p38 inhibition) |
80% KO-DMEM, 2 mMol L-glutamine,10ng/ml bFGF, 1.2 mMol B-ME, 7.5% FCS | 4.5mM | 50% beating CM | (219) (220) |
| EB-Method hESCs |
Guided differentiation: h-Activin A, h-BMP4, h-bFGF, h-VEGF, h-DKK1 |
StemPro-34 (Base) + [L-glutamine, AA, optional MTG, P/S] | >25mM | KDR+ selected cells 35 ± 6% cTNT+, enriched to 57 ± 4% by monolayer culture. DKK1 on day 4 gave 2-fold enrichment of CTNT+ cells | (221) |
| KDR+/PDGFRA+ cells, 50–70% cTNT+ cells in beating EBs. 80% cTNT+ cells by monolayer culture |
(203) | ||||
| EB-method iPSCs |
Forced aggregation EBs with guided differentiation:BMP4, FGF2, Staged O2 levels | RPMI (L-glutamine) 20% FBS on day 3 Insulin on days 0-2 and day 4 |
9–11mM Note: 2uM FA |
Contracting EBs contained 64–89% of cardiac troponin I+cells | (222) |
| Monolayer hiPSCs |
Guided differentiation: h-Activin A, h-BMP4, h-bFGF | StemPro-34 + [L-glutamine,MTG, AA, P/S] | >25mM | Spontaneously beating sheets of CMs 40 ± 15% CMs | (30) |
| Monolayer hESCs hiPSCs |
Guided differentiation method: h-Activin A, h-BMP4, h-bFGF, + Matrigel or CHIR99021, IWP4/IWP2 |
RPMI+B27(-insulin) [d0-6 of differentiation]; RPMI+B27 [from d7 of differentiation] |
11.1mM Note: 2uM oleic acid |
Matrigel addition on day−2 and day 0 generated 80% cTnT+ CMs | (223) |
| Spontaneously beating sheets of CMs, 87% cTNT+ CMs | (207) (206) (205) |
B-ME, beta-mercaptoethanol; CM, cardiomyocytes; MTG, monothioglycerol; P/S, penicillin/streptomycin; AA, ascorbic acid; NEAA, non-essential amino acids.