Cell type Protocol
Duration
Phase I - Endoderm induction factors Phase II - Hepatic induction factors Phase III - Hepatic maturation factors Reference
iPSCs 12 days Day 1-3;
activin A (100 ng/mL),
Wnt 3a (50 ng/mL), HGF (10 ng/mL).
Day 4-7; L-glutamine (1 mM), NAA (1%),  2-Mercaptoethanol (0.1 mM), DMSO (1%). Day 8-12;
OSM (20 ng/mL),
Dexamethasone (0.5 μM), ITS premix (50 mg/mL).
Chen et al. [49]
iPSCs 25 days Day 1-3;
CHIR99021 (3 μM),
Ly294002 (10 μM),
Activin (100 ng/ml),
FGF2 (40 ng/ml),
BMP4 (10 ng/ml).
Day 4-8;
Activin A (100 ng/ml), FGF2 (40 ng/ml),
and B27.
Day 9-25;
HGF (20 μg/ml),
OSM (10 μg/ml).
Rashid et al. [36]
hESCs 20 days Day 1-5;
Activin (100 ng/mL), 
FGF2 (20 ng/mL), 
BMP4 (10 ng/mL), 
LY294003 (10  μM).
Day 6-10;
FGF10 (50 ng/mL), Retinoic acid (10 7 M),  SB431542 (10  μM).
Day 11-20;
FGF4 30 ng/ml, 
HGF 50 ng/mL, 
EGF 50 ng/Ml.
Touboul et al. [46]
hESCs 18 days Day 1-3;
Activin A (100 ng/ml), ITS (1%).
Day 4-8;
FGF4 (30 ng/ml), 
BMP2 (20 ng/ml).
Day 9-18; HGF (20 ng/ml) for 5 days, subsequently OSM (10 ng/ml),
dexamethasone (0.1 μM).
Cai et al.[41]
hESCs 14 days Day 1-3;
Activin A (100ng/mL),
FGF-2 (100ng/ml).
Day 4-11; 10% FBS or KSR, NAA (1 mM) L-glutamine, 1% DMSO, HGF (100 ng/mL). Day 11-14;  10% FBS or KSR, NAA (1 mM),
L-glutamine, dexamethasone (10−7 M).
Basma et al. [42]
hESCs or iPSCs 21 days Day 1-4;
Activin-A (100 ng/ml),
b-FGF (8 ng/ml),
Wnt-3A (25 ng/ ml).
Day 5- 10;
FGF-10 (50 ng/ml),
Retinoic acid (0.1 mM),
SB431542 (1 mM).
Day 11-21;
FGF-4 (30 ng/ml),
EGF (50 ng/ml),
HGF (50 ng/ml).
Wu et al.[75]
HGF: hepatocyte growth factor; NAA: non-essential amino acids; DMSO: dimethyl sulfoxide; OSM: Oncostatin M; ITS: insulin-transferrin-selenium; FGF: fibroblast growth factor; BMP: bone morphogenetic protein; EGF: epidermal growth factor; FBS: fetal bovine serum; KSR: knockout serum
Table 1: List of various methods to differentiate pluripotent stem cells to mature hepatic lineage cells.