| DMEM/F12 AdDMEM/F12 (Invitrogen) |
DMEM/F12 |
low concentration basic fibroblast growth factor (4 ng ml−1) |
| 1% N2 and 1% B27 (GIBCO) |
penicillin and streptomycin |
50 μM Rho-associated protein kinase (ROCK) inhibitor49 (Calbiochem) |
| 1.25 mM N-Acetylcysteine (Sigma) |
10 mM HEPES |
100 N2 supplement (Invitrogen) |
| 10 nM gastrin (Sigma) |
Glutamax N2, B27 (all from Invitrogen) |
Glutamax (Invitrogen) |
| 50 ng/ml EGF (Peprotech) |
1 μM N-acetylcysteine (Sigma) |
minimum essential media-nonessential amino acids (MEM-NEAA) |
| 10% RSPO1 conditioned media (homemade) |
50 ng ml−1 mouse epidermal growth factor (mEGF) |
1 μg ml−1 heparin50 (Sigma) |
| 100 ng/ml FGF10 (Peprotech) |
50% Wnt3a-conditioned medium (WCM) |
1:1 mixture of DMEM/F12 |
| 25 ng/ml HGF (Peprotech) |
10% noggin-conditioned medium (NCM) |
1:200 N2 supplement (Invitrogen) |
| 10 mM Nicotinamide (Sigma) |
20% Rspo1-conditioned medium |
1:100 B27 supplement without vitamin A (Invitrogen) |
| 5 uM A83.01 (Tocris) |
10 μM nicotinamide (Sigma) |
3.5 μl l−1 2-mercaptoethanol |
| 10 uM FSK (Tocris) |
10 nM gastrin (Sigma) |
1:4,000 insulin (Sigma) |
| Noggin (Peprotech) |
500 nM A83-01 (Tocris) |
1:100 Glutamax (Invitrogen) |
| 30% Wnt CM (homemade) |
10 μM SB202190 (Sigma) |
1:200 MEM-NEAA |
| 10 uM (Y27632, Sigma Aldrich) |
Matrigel |
B27 supplement with vitamin A (Invitrogen) |
| Matrigel |
|
Matrigel |
