REAGENT or RESOURCE	SOURCE	IDENTIFIER
Chemicals, peptides, and recombinant proteins
Geltrex LDEV-free, hESC-qualified, reduced growth factor basement membrane matrix	Thermo Fisher Scientific	Cat#A1413301
Recombinant human FGF-2	PeproTech	Cat# 100-18B
Recombinant human Noggin	PeproTech	Cat# 120-10C
Recombinant human BMP-4 (E. coli derived)	PeproTech	Cat#120-05ET
PD 173074	Sigma-Aldrich	Cat#P2499
A 83-01	Tocris Bioscience	Cat#2939
NaHCO3	Sigma-Aldrich	Cat#S5761
Cultrex 3D Culture Matrix Rat Collagen I	Bio-Techne	Cat#3447-020-01
Fluorescein isothiocyanate-dextran, 10 kDa	Sigma-Aldrich	Cat#FD10S
Tetramethylrhodamine isothiocyanate-dextran, 155 kDa	Sigma-Aldrich	Cat#T1287
TrypLE Select (1X)	Thermo Fisher Scientific	Cat#12563011
ROCK inhibitor (Y-27632)	Sigma-Aldrich	Cat#SCM075
Hank’s balanced salt solution (HBSS)	Sigma-Aldrich	Cat#H6648
HEPES, 1 M buffer solution	Thermo Fisher Scientific	Cat#15630049
FBS	Gibco/ATCC	Cat#A13450
Critical commercial assays
TeSR1 medium	In-house. Ludwig et al.3; also available from STEMCELL Technologies	N/A
Buffer RLT	QIAGEN	Cat#79216
Cell lysis buffer (10X)	Cell Signaling Technology	Cat#9803
Halt protease and phosphatase inhibitor cocktail (100X)	Thermo Fisher Scientific	Cat#78440
Experimental models: Cell lines
ChiPS4 hiPSC cell line	Cellartis	CVCL_RM97
Software and algorithms
Prism 9	GraphPad	https://www.graphpad.com/scientific-software/prism/
Fiji (ImageJ)	ImageJ	https://imagej.nih.gov/ij/download.html
Other
OrganoPlate 3-lane 40	MIMETAS	Cat#4003-400B
OrganoFlow	MIMETAS	Cat#MI-OFPR-L
Incucyte S3 live-cell analysis instrument	Sartorius	N/A
ErgoOne E single-channel pipette electronic, 0.5–10 μL	Starlab	Cat#G9001-0010
ErgoOne E 8-channel electronic pipette, 15–300 μL	Starlab	Cat#G9008-0300
Sorvall ST 8 small benchtop centrifuge	Thermo Fisher Scientific	Cat#75007200
Centrifuge 5425 R	Eppendorf	Cat#5425 R
Galaxy Mini centrifuge	VWR	C1413-230 EU