Table 2.

In vitro protocols of MSCs isolation and differentiation in- to-hepatocyte-like cells

Isolation of MSCs	Induction growth factors	Methods of hepatic differentiation	Culture (2D/3D)	References
Enzymatic	Animal model In vivo differentiation	Animal model In vivo differentiation	3D In-vivo	Chen et al. 201612
Result :Transplanted hUCMSCs into hepatectomized rats differentiated into hepatocyte-like cells
Enzymatic	EGF and bFGF, HGF, bFGF and nicotinamide, Dex, ITS premix and OSM	3 steps protocol	2D	Lee, et al. 201213
Result :Potential for hepatogenic differentiation of placenta-derived stem cells (PDSCs) containing WJ
Explant	HGF,FGF-4,OSM, Dex 2	2 steps protocol	2D	Esmaeli 201415
Result: Shift in the fatty acid profile during the hepatocyte differentiation
Enzymatic	EGF, bFGF (2 days) HGF, bFGF (10 days) nicotinamide, ITS (10 days)	Sequential 3 steps	2D	Campard, et al. 200831
Result: Human MSCs can be considered as a source with hepatogenic potential
Explant	bFGF, EGF,	Cocktail	3D	Su et al. 201432
Result: Poly (3-hydroxybutyrate-co-3-hydroxyvalerate-co-3-hydroxyhexanoate) scaffolds loaded with UC-MSCs or differentiated UC-MSCs promoted the recovery of injured livers
Enzymatic	DMEM/F-12 medium with HGF, EGF, ITS, Dex, OSM	2 steps protocol	2D	Zhou et al. 201457
Result: Stimulation of host hepatocyte regeneration via hUCMSCs
Explant	40 ng/mL HGF And 10 ng/mL FGF-4	Cocktail	2D	Zhang et al. 200958
Result: UC-MSC can differentiate into functional hepatocyte-like cells and have advantages over BM-MSC
Explant	EGF and FGF4, HGF, FGF4, Dex, ITS and OSM	2 steps protocol	3D	Chitrangi et al. 201759
Result: gelatin-vinyl acetate scaffold enhanced differentiation of UC-MSCs to hepatocyte-like cells
Ex-vivo	-	Ex vivo	-	Gomez-Aristizabl and Davies 201260
Result: Human UC perivascular Cells (hucpvcs) can act as stromal cells for rat hepatocytes,
Explant	10 ng/mL HGF, 10 ng/mL FGF4	Cocktail	2D	Ewida, et al. 201661
Result: hMSCs can differentiate in vitro into functional hLCs (hepatocyte-like stem cells) in a liver fibrosis rat model
Explant	Liver homogenate supernatants (LHS)	Cocktail	2D	Xue, et al. 201662
Result: Liver tissue microenvironment may contribute to the differentiation of hUCMSCs into hepatocytes both in vitro and in vivo
Enzymatic	Dex, EGF, HGF, ITS and OSM	2-step (for 2 weeks)	2D	Liang, et al. 201263
Result: Extending hUCMSCs replicative lifespan without influencing hepatogenic differentiation potential via telomerization of hUCMSCs by human telomerase reverse transcriptase (HTERT)
Explant	FGF-4 and HGF(14 days) HGF, ITS, oncostatin and Dex. (14 days)	Sequential 2 steps	3D	Raut, and Khanna 201664
Result: improved expression of hepatocyte-specific miRNAs, miR-23b cluster (miR-27b-3p, miR-24-1-5p and miR-23b-3p), miR-30a-5p, miR-26a-5p, miR-148a-3p, miR-192-5p, miR-122-5p due to VPA pre-treatment
Explant	ITS, HGF, OSM, Dex (15 days).	Cocktail	2D	An, et al. 201465
Result: The lineage conversion of hUCMSCs to hepatic cell fate by upregulating the expression of endodermal genes through AKT and ERK activation by valproic acid (VPA)
Explant	EGF, bFGF, HGF, bFGF nicotinamide, OSM, ITS	2 steps protocol	2D	Zheng, et al. 201566
Result: Oncostatin M (OSM) acts an important role in hepatogenic differentiation.
Explant	HGF, infection with a lentivirus containing a miRNA inhibitor sequence	Cocktail	2D	Cui, et al. 201367
Result: The capability of miRNAS to converting hMSCs to a hepatocyte phenotype in vitro
Explant	IGF-I, HGF and Dex OSM up to 23 days	2 steps	3D cell aggregate	Talaei-Khozani, et al. 201568
Result: Facilitating hepatocyte differentiation may be by the UCMSCs aggregate formation before administration of the differentiation protocols

HGF: hepatocyte growth factor; bFGF: basic fibroblast growth factor; OSM: Oncostatin M; Dex: dexamethasone; EGF: epidermal growth factor; ITS: insulin-transferrin-selenium; IGF: insulin-like growth factor; TGF: transforming growth factor; DMEM: Dulbecco’s Modified Eagle’s Medium