REAGENT or RESOURCE	SOURCE	IDENTIFIER
Antibodies
Mouse monoclonal to CD31 (clone WM59), APC	ThermoFisher	Cat. #17-0319-42; RRID: AB_10852842
Mouse monoclonal to CD31 (clone JC/70A)	ThermoFisher	Cat. #MA5-13188; RRID: AB_10982120
Mouse monoclonal to NFATC1 (clone 7A6)	ThermoFisher	Cat. #MA3-024; RRID: AB_2236037
Rabbit monoclonal to VIMENTIN (clone SP20)	ThermoFisher	Cat. #MA5-16409; RRID: AB_2537928
Mouse monoclonal to CD34 (clone 561), PE-Cy7	BioLegend	Cat. #343616; RRID: AB_2629726
Mouse monoclonal to CD56 (clone 5.1H11), APC	BioLegend	Cat. #362504; RRID: AB_2563913
Mouse monoclonal to CD13 (clone WM15), PE	BioLegend	Cat. #301704; RRID: AB_314180
Rabbit monoclonal to NKX2.5 (clone E1Y8H)	Cell Signaling Technology	Cat. #8792; RRID: AB_2797667
Rabbit polyclonal to VE-CADHERIN	Cell Signaling Technology	Cat. #2158; RRID: AB_2077970
Mouse monoclonal to PERIOSTIN (clone F-10)	Santa Cruz Biotechnology	Cat. #sc-398631; RRID: AB_2934053
Rabbit polyclonal to SOX9	MilliporeSigma	Cat. #AB5535; RRID: AB_2239761
Mouse monoclonal to αSMA (Clone 1A4)	Abcam	Cat. #ab7817; RRID: AB_262054
Mouse monoclonal to PDGFRβ (clone 28D4), BV786	BD Biosciences	Cat. #743038; RRID: AB_2741234
Goat anti-Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488	ThermoFisher	Cat. #A-1100; RRID: AB_2534069
Goat anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488	ThermoFisher	Cat. #A-11008; RRID: AB_143165
Goat anti-Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 568	ThermoFisher	Cat. #A-11004; RRID: AB_2534072
Goat anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 568	ThermoFisher	Cat. #A-11011; RRID: AB_143157
Chemicals, peptides, and recombinant proteins
Recombinant Human BMP10	R&D	Cat. #2926-BP
Recombinant Human bFGF	R&D	Cat. #233-FB
Recombinant Human VEGF-165	R&D	Cat. #293-VE
Recombinant Human/Mouse/Rat BMP2	R&D	Cat. #355-BM
Recombinant Human TGF-beta 2	R&D	Cat. #302-B2
Human Apo-Transferrin	R&D	Cat. #3188-AT
Penicillin/streptomycin	ThermoFisher	Cat. #15140122
GlutaMAX	ThermoFisher	Cat. #35050061
Bovine Serum Albumin Fraction V (BSA)	ThermoFisher	Cat. #15260037
Ascorbic Acid	MilliporeSigma	Cat. #A4544
2-Mercaptoethanol	MilliporeSigma	Cat. #M3148
DAPI	MilliporeSigma	Cat. #9564
Fetal Bovine Serum (FBS)	MilliporeSigma	Cat. #F4135
Fibronectin Human Plasma	MilliporeSigma	Cat. #F0895
Triton X-100	MilliporeSigma	Cat. #93418
Sodium Azide	MilliporeSigma	Cat. #S2002
Tween-20	MilliporeSigma	Cat. #P1379
DNase I	MilliporeSigma	Cat. # 260913
DMSO	MilliporeSigma	Cat. #D2650
4% Paraformaldehyde	ThermoFisher	Cat. # J19943K2
CHIR 99021	Tocris	Cat. #4423
Collagenase Type II	Worthington	Cat. #LS004176
Thiazovivin	Selleckchem	Cat. #S1459
Matrigel, Growth Factor Reduced	Corning	Cat. #354230
HEPES	Research Products International	Cat. #H75050
Glycine	Bio-Rad	Cat. #1610718
Proteinase K	Qiagen	Cat. #19131
Accutase	Accutase	Cat. #AT104
Critical commercial assays
Superscript IV VILO	ThermoFisher	Cat. #11756050
PowerTrack SYBR Green	ThermoFisher	Cat. #A46012
RNA 6000 Nano Kit	Agilent	Cat. # 5067-1511
CD31 MicroBead Kit	Miltenyi Biotec	Cat. #130-091-935
CD34 MicroBead Kit	Miltenyi Biotec	Cat. #130-046-702
Deposited data
Raw & processed data from iPSC-derived populations	This study	GEO: GSE226476
15-week-old fetal human valve single-cell RNAseq data	Liu et al., 202353	GEO: GSE228638
Experimental models: Cell lines
WTC11 (ACTN2-eGFP tagged, monoallelic)	Coriell Institute	Cat. #AICS-0075-085
MSN02-452,53	Gift from Dr. Dubois	N/A
Oligonucleotides
See Table S1	This study	Table S1
Software and algorithms
FCS Express 7	De Novo Software	https://denovosoftware.com/
GraphPad Prism	GraphPad	https://www.graphpad.com/
R CRAN, Version 4.2.0		https://cran.r-project.org/
STAR v.2.7.10a54		https://github.com/alexdobin/STAR
Picard v2.26.10		http://broadinstitute.github.io/picard/
HTSeq v2.0.255		https://htseq.readthedocs.io/en/master/
DESeq256		https://bioconductor.org/packages/release/bioc/html/DESeq2.html
Enrichr58		https://maayanlab.cloud/Enrichr/
Seurat v467		https://satijalab.org/seurat/
FIJI		https://imagej.net/software/fiji/
Other
RPMI 1640	ThermoFisher	Cat. #11875093
B27 Supplement	ThermoFisher	Cat. #17504044
B27 Supplement, Minus Insulinf	ThermoFisher	Cat. #A1895601
DMEM, high glucose	ThermoFisher	Cat. #11965092
PBS, no calcium, no magnesium	ThermoFisher	Cat. #14190144
6-well clear flat bottom TC-treated culture plates	Fisher Scientific	Cat. #FB012927
24-well clear flat bottom TC-treated culture plates	Fisher Scientific	Cat. #FB012929
40um cell strainer	Fisher Scientific	Cat. #22363547
Fibroblast Growth Medium, Kit	PromoCell	Cat. #C-23110
Endothelial Cell Growth Medium MV2, Kit	PromoCell	Cat. #C-22121
HBSS	Corning	Cat. #21022CV
EasySep Buffer	StemCell Technologies	Cat. #20144
12-well chamber slide	Ibidi	Cat. #81201
Coverslips	Ibidi	Cat. #10811
VECTASHIELD PLUS Antifade Mounting Medium	Vector Laboratories	Cat. #H-1900
LS Column	Miltenyi Biotec	Cat. #130-042-401
Cell Counting Slides	Bio-Rad	Cat. #1450015
Trypan Blue	Bio-Rad	Cat. #1450021
Human Reference cDNA	Takara	Cat. #636693