Pancreatic tumor organoid growth medium

Reagent	Final concentration	Amount
F12+++	N/A	14 mL
Wnt3a-conditioned medium∗	50%	25 mL
R-spondin-1-conditioned medium∗	10%	5 mL
mNoggin-conditioned medium∗	10%	5 mL
B27 (50×)	1×	1 mL
Nicotinamide (1 M)	10 mM	500 μL
N-Acetylcysteine (0.5 M)	1.25 mM	125 μL
hEGF (200 μg/mL)	50 ng/mL	12.5 μL
hFGF-10 (100 μg/mL)	100 ng/mL	50 μL
hGastrin I (100 μM)	10 nM	5 μL
A83-01 (500 μM)	500 nM	50 μL
Primocin (50 mg/mL)	100 μg/mL	100 μL
Total	N/A	50 mL

Once prepared, store 10 mL aliquots at −20°C for up to 3 months.

∗For production and activity testing of conditioned media, see Farin et al. (2012) and Boj et al. (2017).