Complete media for airway organoid generation (AO)

Reagent	Final concentration	Amount
AdDF+ (See above)	N/A	29.115 mL
R-Spondin 1 Reconstitute in 1× PBS 0.1% BSA at 200 μg/mL	500 ng/mL	75 μL
FGF-7 Reconstitute in 1× PBS 0.1% BSA at 20 μg/mL	25 ng/mL	37.5 μL
FGF-10 Reconstitute in 1× PBS 0.1% BSA at 100 μg/mL	100 ng/mL	30 μL
Noggin Reconstitute in 1× PBS 0.1% BSA at 40 μg/mL	100 ng/mL	75 μL
B-27 supplement (50×)	1×	600 μL
Primocin (50 mg/mL)	50 μg/mL	30 μL
A83-01 Reconstitute in DMSO at 10 M/mL	500 nM/mL	1.5 μL
Y-27632 (10 mM/mL)	5 μM/mL	15 μL
SB202190 Reconstitute in DMSO at 10 mM/mL	5 μM/mL	15 μL
N-Acetylcysteine Reconstitute in DMSO at 12.5 M/mL	1.25 mM/mL	3 μL
Nicotinamide Reconstitute in DMSO at 50 M/mL	5 mM/mL	3 μL
Total	N/A	30 mL

Always prepare and use fresh.