Practical choice for robust and efficient differentiation of human pluripotent stem cells

Table 1 Summary of current approaches for human pluripotent stem cells differential direction into targeted cells or tissues

Targeted cells or tissues	Cultural approaches				Check points of differential status			Ref.
	EB formation	2D or 3D	2D Surface or 3D system	Multiple cells co-culture	Gene expression	Protein level	Under in vitro or in vivo
Neural progenitor	-	2D	Matrigel	-	PAX6, TBR2	-	In vitro	[21]
Brain	EB	3D	Low attachment plate, Matrigel	-	PAX6, SOX2, FOXG1, TBR2, ARHGAP11B	Nestin	In vitro	[21]
Midbrain	EB	3D	Matrigel	-	SOX1, VMAT2, TH, NURR1, DAT, GIRK2, PITX3, AADC, ANLN, FAH, MBP, GLA35ST1, ACSL1, CLDN11, CHAT, MAPT, GFAP, S100B, ALDH1L1	-	In vitro	[30]
Brain	-	3D	Microwell, bioreactor	Neuronal, astrocyte	-	-	In vitro	[54]
Retinae	-	3D	Matrigel	-	SIX3, PAX6, RAX, OTX2, VSX2, PRKCZ, MITF	TJP1, LAMB1, RHO, OPN1LW/OPN1MW, OPN1SW	In vitro	[31]
Retinae	EB	3D	Low attachment plate	-	BRN3B, PAX6, RAX, SIX3, LHX2, CHX10, OTX2	RHO, PKCα, Arl13b, OPN1SW, OPN1MW	In vitro	[37]
Retinae	-	3D	Matrigel	RPE, retinal organoid	CRX, RCVRN, NRL, GNAT1, RHO, CHX10, OPN1LW/MW, OTX2, RLBP1, PROX1	-	In vitro	[32]
Melanocyte	EB	3D	Microwell, low attachment plate	-	MITF, PAX3, SOX10, KIT, TYR, TYRP, DCT, PEML	Melanin	In vivo	[12]
Hepatic stellate cells	-	2D	Matrigel	-	NCAM, KDR, PDGFRα, P75NTR, ALCAM, ACTA2, COL1α1, LRAT, RELN, PCDH7, PDGFRβ, SYP, GFAP, PPARγ, NGF, α-SMA	Desmin, PDGFRβ, P75NTR, ALCAM, PDGFRα, CD73, KDR, NCAM	In vitro	[23]
Liver	EB	3D	Microwell	iPSC endoderm cell, HUVEC, BM-MSC	ALB, G6PC, CYP2C9, CYP2C19, CYP3A4, CYP3A7	CYP3A4, ALB, Urea, NTCP	In vitro	[52]
Liver	-	3D	Microwell	iPSC-tHE, iPSC-EC, iPSC-STM	TBX3, ADRA1B	AFP, ALB, Complement factor H, Coagulation factor VIII, Transferrin, AAT	In vivo	[50]
Intestinal	-	3D	Matrigel	-	KLF5, ECAD, SOX9, KI67	Villin	In vivo	[33]
Entersphere	-	3D	Matrigel	Pan-epithelium cell, HLF, HUVEC	SOX9, CK20, CDX2, NNKX2.1, LGR5, OLFM4, TACSTD2, VIL1, APOA1, FABP2	E-cadherin, Cytokeratin18, α-SMA	In vitro	[34]
Cardiomyocyte	-	2D	Matrigel	-	-	TNNT2, ACTN2	In vitro	[67]
Cardiomyocyte	EB	3D	Low attachment plate	-	TBX5, NKX2.5, GATA4	TNNT2, TNNI3, MYH6, MYL7	In vitro	[47]
Cardiomyocyte	EB	3D	Suspension bioreactor	-	-	TNNT2, α-Actinin, MLC-2v, MLC-2a	In vitro	[13]
Heart	EB	3D	Matrigel	hESC-CPC, hESC-MSC, HUVEC	KDR, MESP1, NKX2.5, TBX5, GATA4, ISL1, PDGFR-α, MEF2C, CD90, CD73, CD105, CD44, CD31, cTNT, β-MHC, MLC2v, KCNA4, KCNJ2, KCNH2	-	In vivo	[35]
Hematopoietic cell	EB	3D	-	-	RUNX1, SCL/TAL1	CD34, CD43, CD45	In vitro	[68]
T Cell	-	2D	-	-	TRA, TRB, RAG1, RAG2	CD8ab, LMP2, TCR, TCRab-CD3	In vivo	[69]
Macrophage	EB	3D	Low attachment plate	-	MAF, CSFR1, FLT3, CCR2	CD14, CD45, CD11b, CD16, TNF-α	In vitro	[11]
Liver sinusoidal endothelial cell	EB	3D	Low-cluster plate	-	CD31, CDH5, CD34, F8, STAB2, LYVE1, FLK1, FLT4, FCGR2B	-	In vitro	[55]
Platelet	-	3D	Ultra-low attachment plate	-	CD41a, CD13, CD42b, CD31, CD34, CD43, CD41b	Thrombospondin4, Platelet factor 4	In vivo	[49]
Mesenchymal cell	-	2D	Matrigel	-	CD146, CD73, CD140a, CD90, CD105, CD44, PDGFRβ, CSPG4, NES, LEPR, ADRB2, KITLG, IGFBP2, TNC, CXCL12, ADRB3	-	In vitro	[26]

-: None; EB: Embryoid body; 2D: Two-dimensional; 3D: Three-dimensional; RPE: Retinal pigment epithelium; iPSCs: Induced pluripotent stem cells; HUVECs: Human umbilical cord vein endothelial cells; BM-MSC: Bone marrow mesenchymal stem cell.