Table 1 Composition of organoid and monolayer culture media reagents
Media reagents | Manufacturer | Final Concentration |
|---|---|---|
Basal Medium | Â | Â |
 Advanced DMEM/F12 | Gibco | NA |
 GlutaMAX Supplement | Gibco | 2 mM |
 HEPES | Gibco | 10 mM |
 Penicillin/streptomycin | Gibco | 1x |
Organoid Expansion Medium (OEM) | Â | Â |
Per Basal Medium including: | Â | Â |
 Noggin conditioned medium - engineered HEK293 cells, Baylor College of Medicine | Lab-prepared | 10% |
 R-spondin conditioned medium - Cultrex HA-R-spondin1-Fc 293T cells, R&D Systems | Lab-prepared | 20% |
 Recombinant murine Wnt-3a | PeproTech | 100 ng/mL |
 A-83-01 | Sigma-Aldrich | 500 nM |
 B27 | Gibco | 1x |
 Murine EGF | R&D Systems | 50 ng/mL |
 Gastrin | Sigma-Aldrich | 10 nM |
 N2 | R&D Systems | 1x |
 Nicotinamide | Sigma-Aldrich | 10 mM |
 N-Acetyl-L-cysteine | MP Biomedicals | 1 mM |
 SB202190 | Sigma-Aldrich | 10 µM |
 Primocin | InvivoGen | 100 µg/mL |
Organoid Differentiation Medium (ODM) | Â | Â |
Per OEM omitting: | Â | Â |
 Noggin conditioned medium |  |  |
 R-spondin conditioned medium |  |  |
 Recombinant murine Wnt-3a |  |  |
For jejunum and duodenum organoids | Â | Â |
Per OEM and ODM including: | Â | Â |
 Fetal bovine serum | Gibco | 10% |
Monolayer Expansion Medium (MEM) | Â | Â |
Per OEM including: | Â | Â |
 Y-27,632 | STEMCELL Technologies | 10 µM |
 CHIR99021 | Sigma-Aldrich | 100 nM |
 LY2157299 | Sigma-Aldrich | 500 nM |
 Fetal bovine serum | Gibco | 20% |
Monolayer Differentiation Medium (MDM) | Â | Â |
Per MEM omitting: | Â | Â |
 Noggin conditioned medium |  |  |
 R-spondin conditioned medium |  |  |
 Recombinant murine Wnt-3a |  |  |