. 2020 Aug 17;26(4):367–382. doi: 10.1089/ten.teb.2020.0091

Table 2.

Summary of Differentiation Protocols for Human Pluripotent Stem Cell-Derived Pericytes

Cell source	Differentiation methods	Characterizations	References
ESI-017 hESCs	Spontaneous EB differentiation; EC medium	CD146, CD105, PDGFRβ	Greenwood-Goodwin et al.¹⁰²
H9, I6 hESCs	Spontaneous EB differentiation	CD105, NG2, PDGFRβ, calponin, nestin	Dar et al.¹⁰³
C3, KTR13 hiPSCs	Spontaneous EB differentiation	CD105, NG2, PDGFRβ, calponin, nestin	Dar et al.¹⁰³
NL-HES4 hESCs	Mesoderm induction: BMP4, Activin A, VEGF	CD105, NG2, PDGFRβ, CD146, CD73, CD44, 30% yield; functional in cultured vascular plexus and zebrafish xenografts	Orlova et al.; Iendaltseva et al.^106,120,139
hiPSCs	Mesoderm induction: BMP4, Activin A, VEGF		Orlova et al.; Iendaltseva et al.^106,120,139
H9 hESCs	Mesoderm induction: 10% serum in α-MEM; then VEGF and SB431542 in EC medium	NG2, PDGFRβ, CD73, CD44; studied the differences compared to vSMCs (SMA, SMMHC, calponin). ECM protein production, collagens and elastin, MMP2, 9, 14, migration and invasion	Wanjare et al.¹⁰⁸ with Jamieson et al.⁴¹
hiPSCs			Wanjare et al.¹⁰⁸ with Jamieson et al.⁴¹
H1 hESCs	Mesoderm induction: BMP4, VEGF, FGF-2 in XVIVO-15 medium; later using SB431542; CD325-CD56+ cells were sorted using FACS	The derived cells coexpressed common mesenchymal markers (CD73, CD105, CD90, and PDGFRβ) with a subset of cells defined as CD146^hiCD73^hi. Transcriptional similarity exists between hPSC-derived CD146⁺⁺ and primary human CD146⁺⁺ perivascular cells	Chin et al.¹¹¹
UCLA3 and UCLA6 hESCs			Chin et al.¹¹¹
H1, H9 hESCs	Mesoderm induction: coculture with OP9 stromal layer; or with BMP4, Activin A, and FGF-2; with BMP4, CHIR99021, PDGF-BB, and Activin A	MBs differentiate into primitive PDGFRβ⁺CD271⁺CD73⁻ mesenchymal progenitors, which give rise to proliferative PCs, SMCs, and MSCs. MB-derived PCs can be further specified to CD274⁺ capillary and DLK1⁺ arteriolar PCs	Kumar et al.¹¹⁰ Blanchard et al.¹³⁸
hiPSCs			Kumar et al.¹¹⁰ Blanchard et al.¹³⁸
H9 hESCs	Mesoderm induction: MIM-mPCs	NG2, PDGFRβ, CD13, CD146; brain specific markers VTN, FOXC1, FOXF2. FOXF2 is higher in ncPCs than mPCs Functional assay: coculture with ECs forms tube-like structures; Wnt inhibition by DKK1 reduced PAX7 expression	Faal et al.¹¹⁵
hiPSCs from AD patients, lines AD 5, 6 (APOE4), 13, 14, 20, 22 (APOE3), 29	Neural crest induction: Wnt activation using CHIR99021, DMEM/F12 plus B27-ncPCs		Faal et al.¹¹⁵
H9 hESCs	Neural crest induction: Wnt activation using CHIR99021, plus SB431542, FGF2, dorsomorphin, heparin in E6 medium; use MACS sorting and 10% FBS	PDGFRβ, NG2, expressing CNN1, TAGLN, ANPEP, TBX18; brain mural markers: FOXF2, ZIC1, ABCC9, KCNJ8; functional assay: the derived cells assemble with vascular cord networks; induced BBB properties	Stebbins et al.¹¹⁸
IMR90C4 and CS03n2 iPSCs			Stebbins et al.¹¹⁸

DKK1, Dickkopf WNT signaling pathway inhibitor 1; FBS, fetal bovine serum; MACS, magnetic-activated cell sorting; SMMHC, smooth muscle myosin heavy chain; PC, pericyte; BBB, blood–brain barrier; hiPSC, human induced pluripotent stem cell; hESC, human embryonic stem cell; FGF, fibroblast growth factor; MIM, mesodermal induction medium; BMP4, bone morphogenetic protein 4; PDGF-B, platelet-derived growth factor-B; FACS, fluorescence-activated cell sorting; VEGF, vascular endothelial growth factor; EC, endothelial cell; EB, embryoid body; PDGFRβ, platelet-derived growth factor receptor β; NG2, neural/glial antigen 2; vSMC, vascular smooth muscle cell; SMA, smooth muscle actin; hPSC, human pluripotent stem cell; MB, mesenchymoangioblast; MSC, mesenchymal stem cell; α-MEM, alpha-minimal essential medium; DMEM, Dulbecco's modified Eagle's medium.