Derivation and applications of human hepatocyte-like cells

Table 1 Summary of hepatocyte-like cells differentiation protocols

Ref.	EB / monolayer	Protocol features		In vitro functional assays	In vivo assay
		Endoderm induction	Hepatic specification and maturation
Cai et al[18], 2007	Monolayer	Activin A, ITS	FGF4, BMP2, HGF, OSM, Dex	ALB, Glycogen, ICG, LDL, CYP450	Yes
Hay et al[19], 2008	Monolayer	Activin A, Wnt3a	Serum, DMSO, Insulin, HGF, OSM	Urea, Gluconeogenesis, AFP	Yes
Agarwal et al[24], 2008	Monolayer	Activin A, low serum	FGF4, HGF, OSM, Dex	ALB, Glycogen, ICG	Yes
Basma et al[7], 2009	EB/monolayer	Activin A, bFGF	FGF, DMSO, Dex	ALB, Urea, AAT, CYP450	Yes
Song et al[20], 2009	Monolayer	Activin A	FGF4, BMP2, HGF, KGF, OSM, Dex	ALB, Urea, Glycogen, CYP450	No
Si-Tayeb et al[21], 2010	Monolayer	Activin A	BMP4 FGF2, HGF, OSM	Glycogen, LDL, oil red O storage, ICG, Urea	Yes
Sullivan et al[22], 2010	Monolayer	Activin A, Wnt3a	β-ME, DMSO, Insulin, HGF, OSM	CYP450, Fibrinogen, Fibronectin, Transthyretin, AFP	No
Touboul et al[23], 2010	Monolayer	Activin A, FGF2, BMP4, LY294002	FGF10, RA, SB431542, FGF4, HGF, EGF	Glycogen, CYP450, ICG, LDL	Yes
Borowiak et al[35], 2009	Monolayer	Activin A, Wnt3a, HGF	OSM, Dex, ITS	CYP450, Urea, LDL, Glycogen	Yes
Ogawa et al[34], 2013	EB/monolayer	BMP4, Activin A, Wnt3a	FGF10, bFGF, BMP4, HGF, OSM, Dex, cAMP	ICG, Glycogen, ALB, CYP450	No
Siller et al[37], 2015	Monolayer	CHIR99021	DMSO, dihexa, Dex	ALB, Glycogen, ICG, CYP450	No

EB: Embryoid bodies; KGF: Keratinocyte growth factor; β-ME: 2-mercaptoethanol; RA: Retinoic acid; EGF: Epidermal growth factor; ITS: Insulin, transferrin, selenium; dihexa: Hepatocyte growth factor receptor agonist N-hexanoic-Tyr, Ile-(6) aminohexanoic amide; ALB: Albumin secretion; AFP: Alpha-fetoprotein secretion; AAT: Alpha-1-antitrypsin secretion; LDL: Low-density lipoprotein uptake; glycogen: Glycogen storage; ICG: Indocyanine green uptake; Urea: Urea secretion and production; CYP450: CYP450 activity.

Table 2 Summary of transplantation studies using hepatocyte-like cells

Ref.	Animal model	Route	Proliferative stimulus	Type and number of cells	Donor / recipient (% engrafted)	Donor / recipient (% repopulated)	Time post-transplantation
Agarwal et al[24], 2008	NOD-SCID mice	Portal vein	CCl4-injured	106 hES-DEs	< 1%	NA	28 d
Basma et al[7], 2009	NOD-SCID mice	Spleen	Retrorsine and partial hepatectomy	1 × 106 hES-HLCs	NA	NA	21 d
Liu et al[77], 2011	NSG mice	Tail vein	dimethylnitrosamine -injured	0.1 - 2 × 106 hiPSC- multistage hepatic cells	2%–17%	8%–15%	56 d
Asgari et al[76], 2013	Normal mouse	Tail vein	CCl4-injured	1 × 106 hiPSC- HLCs	2 ± 0.7%	NA	35 d
Carpentier et al[73], 2014	MUP-uPA/ SCID/Bg mice	Spleen	NA	4 × 106 hiPSC-HLCs	1%-7%	< 1 to up to 20%	100 d
Song et al[20], 2015	Immunocompetent mice	Intraperitoneal cavity	NA	4.4×105 hiPSC-HLCs in capsules	NA	NA	24 d
Nagamoto et al[74], 2015	uPA/SCID mice	Spleen	NA	1 × 106 Ad-FNK-transduced hiPSC-HLCs	NA	NA	28 d
Nagamoto et al[78], 2016	Mice	hiPS-HLC sheet transplantation	2/3 partial hepatectomy and CCl4-injured	8 × 105 hiPSC-HLCs	NA	NA	14 d

DE: Definitive endoderm; HLCs: Hepatocyte-like cells; hPSCs: Human pluripotent stem cells.