|
L-WRN cell culture medium
|
|
|
|
| Advanced DMEM/F12 |
|
Life Technologies |
12634–010 |
| Fetal Bovine Serum (FBS) |
1% |
Life Technologies |
10437–028 |
| Penicillin-Streptomycin |
100 U/mL |
Life Technologies |
15140–122 |
|
Washing buffer
|
|
|
|
| Phosphate Buffered Saline (PBS) |
50 mL |
Life Technologies |
10010–023 |
| Penicillin-Streptomycin |
125 U/mL |
Life Technologies |
15140–122 |
| Amphotericin B |
6.25 μg/mL |
Life Technologies |
15290–018 |
|
Organoid culture medium
|
|
|
|
| L-WRN Conditioned medium |
1:1 |
ATCC |
CRL-3276 |
| Advanced DMEM/F12 |
1:1 |
Life Technologies |
12634–010 |
| Penicillin-Streptomycin |
100 U/mL |
Life Technologies |
15140–122 |
| N-Glutamine |
10 μl/mL |
Life Technologies |
35050–061 |
| N-2-hydroxyethylpiperazine-N-2-ethane sulfonic acid, HEPES |
10 mM |
Life Technologies |
15630–080 |
| B27 |
10 μl/mL |
Gibco |
17504–044 |
| N2 |
10 μl/mL |
Gibco |
17502–048 |
|
Organoid seeding medium
|
|
|
|
| Organoid culture medium |
|
|
|
| Epidermal growth factor (EGF) |
50 ng/mL |
Invitrogen |
PMG8041 |
| Fibroblast growth factor-10 (FGF10) |
100 ng/mL |
PeproTech |
100–26 |
|
Primary antibodies
|
|
|
|
| Anti-Cytokeratin 19 (CK19) antibody, Rabbit |
1:250 |
Abcam |
ab53119 |
| Sex-Determining Region Y-Box 9 (SOX9) antibody, Rabbit |
1:200 |
Santa Cruz |
sc-20095 |
| Pancreatic Duodenal Homeobox 1 (PDX1) antibody, Rabbit |
1:2000 |
DSRB |
F109-D12 |
| E-cadherin antibody, Goat |
1:500 |
Santa Cruz |
sc-31020 |
| Acetylated α-tubulin antibody, Mouse |
1:500 |
Sigma-Aldrich |
T6793 |
| Secondary antibodies |
|
|
|
| 488 labeled anti-rabbit, Donkey IgG |
1:1000 |
Invitrogen |
A-21206 |
| 594 labeled anti-goat, Donkey IgG |
1:1000 |
Invitrogen |
A-11058 |
| 568 labeled anti-mouse, Goat IgG2b |
1:500 |
Invitrogen |
A-21144 |
| TopFlash Wnt reporter assay |
|
|
|
|
TopFlash HEK293 cell line
|
|
ATCC |
CRL-1573 |
| Luciferase Assay Kit |
|
Biotium |
30003–2 |
| 0.05% Trypsin-EDTA |
|
Life Technologies |
25300054 |
| 0.4% Trypan Blue Solution |
|
Life Technologies |
15250061 |
|
Additional materials and reagents
|
|
|
|
| Basement matrix, phenol free Matrigel |
|
CORNING |
356237 |
| Dissociation buffer, Accutase |
|
Gibco |
A1110501 |
| Cell culture freezing medium, Recovery |
|
Life Technologies |
12648010 |
| Cell strainer (70 μm, steriled) |
|
Fisherbrand |
22363548 |
| Guanidinium thiocyanate-phenol RNA extraction, TRIzol |
|
Invitrogen |
15596026 |
| Specimen processing gel, HistoGel |
|
Thermo Fisher Scientific |
HG-4000–012 |
| Universal mycoplasma detection kit |
|
ATCC |
30–1012K |
| 1.5 mL microcentrifuge tube |
|
Fisherbrand |
05–408–129 |
| 24 well plate |
|
USA Scientific |
CC7682–7524 |
| 50 mL conical centrifuge tube |
|
Fisher scientific |
14–432–22 |
| Fluorescence microscope |
|
Nikon |
Eclipse E800 |
| Inverted microscope |
|
Biotium |
30003–2 |
| Necropsy tray |
|
Fisherbrand |
13–814–61 |
