Table 3.

EAC Organoid Medium (50 mL)

Reagent	Volume	Final concentration
Basal Medium	24 mL
WRN Conditioned Medium	24 mL	50%
N-2 (100x)	500 μL	1x
B-27 (50x)	1 mL	1x
NAC (0.5 M)	100 μL	1 mM
CHIR99021 (5 mM)	5 μL	0.5 μM
EGF (500 ng/μL)	25 μL	250 ng/mL
A83–01 (5 mM)	5 μL	0.5 μM
SB202190 (10 mM)	5 μL	1 μM
Gastrin (1 mM)	5 μL	0.1 μM
Nicotinamide (1 M)	1 mL	20 mM
Y-27632 (10 mM)	50 μL	10 μM
Gentamicin (50 mg/mL)	5 μL	10 μM§
Antibiotic-Antimycotic (x100)	500 μL	1x
FGF-10* (100 μg/mL)	50 μL

^§, Note that Basal Medium contains 5 μM Gentamicin before this supplementation.

^*, only added when establishing primary cultures and recovering from frozen stocks.