Table 2 Summary of culture medium compositions for liver organoids
From: Liver organoids: an in vitro 3D model for liver cancer study
| Â | Function | Concentrations | |||
|---|---|---|---|---|---|
Human | Mouse | ||||
Healthy | Cancer | Healthy | Cancer | ||
Basal medium | |||||
 Advanced DMEM/F12 | Providing basal nutrition |  |  |  |  |
 HEPES | Maintaining the osmotic pressure | 10 mM | 10 mM | 10 mM | 10 mM |
 B27/N2 | Suppressing cell differentiation, supporting the growth | 1–2% B27 + 1% N2 | 2% B27 + 1% N2 | 2% B27 | 2% B27 + 1% N2 |
 GlutaMAX | Common substitution of l-glutamine | 1% | 1% | 1% | 1% |
 N-Acetyl-l-cysteine | Regulate cell proliferation, differentiation and apoptosis | 1–1.25 mM | 1.25 mM | 1.25 μM/1–1.25 mM | 1.25 μM |
 Nicotinamide | Playing important role in self-renewal of HCC stem cells and cellular energy metabolism | 10 mM | 10 mM | 10 mM | 10 mM |
Cytokines | |||||
 Wnt3a | Ligand of canonical Wnt/β-catenin pathway, supporting strongly increased self-renewal of organ and embryonic stem cells | 800 ng/ml Wnt3a liposomes or 30% Wnt3a—condition medium | 30% Wnt3a—condition medium | 100 ng/ml or 30% Wnt3a—condition medium | 30% Wnt3a—condition medium for initial 3 days |
 BMP7 | Regulating cell proliferation, differentiation, migration, organization, and apoptosis | 25 ng/ml in DM | 25 ng/ml in DM |  |  |
 EGF | Growth factor | 50 ng/ml | 50 ng/ml | 50 ng/ml | 50 ng/ml |
 FGF | Growth factor | 100 ng/ml FGF10 ± 100 ng/ml FGF7 (100 ng/ml FGF19 in DM) | 100 ng/ml FGF10 (100 ng/ml FGF19 in DM) | 50–100 ng/ml FGF10 ± 50 ng/ml FGF7 | 100 ng/ml |
 HGF | Growth factor | 25 ng/ml | 25 ng/ml | 25/40/50 ng/ml | 50 ng/ml |
 Gastrin | Growth factor for colorectal, stomach, liver, and pancreatic cancer | 10 nM | 10 nM | 10 nM | 10 nM |
Inhibitors | |||||
 Noggin | inhibitor of BMP-4 and BMP-7; to inhibit the differentiation of stem cells | 25 ng/ml in IM | 25 ng/ml in IM | 100 ng/ml | 10% Noggin-conditioned medium for initial 3 days |
 A83-01 | ALK4/5/7 inhibitor; to decrease cell motility, adhesion and invasion increased by TGF-β1 | 2-5 μM (0.5 µM in DM) | 5 μM (0.5 µM in DM) | 1 μM (50 nM in DM) |  |
 Y-27632 | Rock inhibitor; to facilitate the attachment of primary cells in vitro, inhibit the apoptosis of embryonic stem cells and promote the self-renewal and proliferation of stem cells | 10 μM | 10 μM | 10–20 μM |  |
 CHIR99021 | GSK3β inhibitor; to enhance Wnt target gene expression | 3 μM |  | 3 μM |  |
Activators | |||||
 R-spondin-1 | Wnt pathway activator | 1 μg/ml or 10–15% RSPO1—condition medium | 10% RSPO1—condition medium | 0.5–1 μg/ml or 15% RSPO1—condition medium | 10% RSPO1—condition medium |
 Forskolin | Adenyl cyclase activator (cAMP activator) | 10 μM | 10 μM |  |  |
References | Â | [36] | |||