TABLE 2.

Protocols and composition of media used by different studies to generate hepatocytes and insulin producing pancreatic β-cells from DPSCs.

Differentiation	Inducers	Confirmation markers and tests	Protocol timeline (days)	References
Hepatogenesis	• Specification: 2%FBS, HGF • Maturation: ITS-X, dex, and OSM	α-fetoprotein, albumin, hepatic HNF-4, IGF-1, glycogen storage, carbamoyl phosphate synthetase, glucagon, and urea secretion	22–28	Ishkitiev et al., 2010
	• Specification: ITS-X and embryo-trophic factor • Differentiation: ITS-X, embryo-trophic factor, and HGF • Maturation: ITS-X, embryo-trophic factor, HGF, dex, and OSM	α-fetoprotein, albumin, hepatic HNF-4, IGF-1 gene expression. Glycogen storage. Carbamoyl phosphate synthetase, glucagon, and urea secretion	22–28	Ishkitiev et al., 2012
	1% FCS; HGF, oncostatin, nicotinamide, LDL, FGF-4, insulin, glucose, and linoleic acid	Mdr-1, cyp-2e1, Erythropoietin, cytokeratin 8, cytokeratin 18, and cytokeratin 19 gene expression. Albumin secretion	40	Ferro et al., 2012
	• Specification: HGF, EGF, and dex • Maturation: ITS, OSM, and dex	Albumin, TAT, and α-fetoprotein secretion	28	Kumar et al., 2017
	• Definitive endoderm generation: KOSR and Act A • Specification: KOSR, FGF-4, and HGF • Differentiation: FGF-4, HGF, OSM, and dex	AAT, G6P, HNF6, and cytokeratin 18 gene expression. CYP3A4 activity and AST activity. Albumin secretion and Glucagon storage	22	Gil-Recio et al., 2020
Pancreatic β-cell like	• Induction: ITS, Act A, sodium butyrate, and β-ME • Differentiation: ITS, taurine, GLP-1, nicotinamide, and NEAAs	C-peptide, Pdx-1, Pax4, Pax6, Ngn3, and Isl-1 gene expression. Insulin and c-peptide secretion	10	Govindasamy et al., 2011; Sawangmake et al., 2014; Kanafi et al., 2013; Matei et al., 2017; Yagi Mendoza et al., 2018
	• Pre-induction: FBS, RA, and β-ME • Induction: nicotinamide, RA, and β-ME. Alternation between L-DMEM and H-DMEM media, each supplemented with FBS, NNAA, zinc sulfate, and selenium	PDX-1, insulin and GLUT-2 gene expression. Dithizone staining. Insulin secretion	21	Carnevale et al., 2013
	• Induction: HGF and αFGF • Differentiation: HGF, αFGF, EGF, and β-ME. Maturation: HGF, nicotinamide, and embryo-trophic factor	Generation of pancreatic -a, -b, -d, and pancreatic polypeptide-producing cells. Expression of Pdx-1Hhex, Mnx1, Neurog3, Pax4, Pax6, Nkx6-1, Ins, GLG, PPY, SST, Glut-2 and AMY2A, and insulin	17	Ishkitiev et al., 2013
	Matrigel (3D biomaterial matrix) • Stage 1: Act A, Noggin, LiCl, polyvinyl chloride, and β-ME • Stage 2: retinoic acid, A83-01, LDE225, A83-01, and polyvinyl chloride • Stage 3: polyvinyl chloride, ITS, SB, nicotinamide, and NNAAs	Gene expression for FOXA1, Sox17, glucagon, Pax4, Insulin, Nkx6-1, Neurog3, NeuroD1, Pdx1, and CXCR4. Immunohistochemistry for Nkx6-1, Pdx-1, CXCR4, Sox17, and insulin	Not mentioned	Xu et al., 2019