Table 1.
Summary of culture medium components for generation of liver tissue-derived liver organoids.
| Function of supplemented components |
liver organoids | liver cancer organoid | |||||||
|---|---|---|---|---|---|---|---|---|---|
| References | Huch et al. (17) | Hu et al. (21) | Hu et al. (21) | Lam et al. (30) | Broutier et al. (23) | Cao et al. (28) | Wang et al. (29) | ||
| Source cells or tissues | Liver ductal cells | Fetal hepatocytes | primary mouse hepatocytes | Nontumoral human liver tissues at the resection margin of surgical HCC | Liver tumor specimens through resection | Primary mouse liver cancer tissues | Hepatobiliary tumor specimens through resection | ||
| Digestion | collagenase-accutase | Type IV collagenase | Type IV collagenase | collagenase type II | collagenase-accutase | 0.5 mg/ml Collagenase type XI; 0.2 mg/ml Dispase; 1% FBS in DMEM | collagenase D ; 0.1 mg mL−1 DNase I ; 2 × 10−6 m Y27632; 100 µg mL−1 Primocin | ||
| Basement membrane simulation | Matrigel or reduced growth factor BME 2 | Matrigel | Matrigel | Matrigel | BME2 | Matrigel | Matrigel | ||
| Culture media | AdDMEM/F12 | Provision of basal nutrition | √ | √ | √ | √ | √ | √ | |
| AIM-V | Provision of basal nutrition without serum | √ | |||||||
| Penicillin-Streptomycin | Antibiotics to prevent of microorganism contamination | √ | √ | √(0.35 U/ml ) | √(1%) | ||||
| L-Glutamine | Essential amino acid for cell growth | √ | |||||||
| Glutamax | Substitution of L-Glutamine | √ | √ | √(1%) | √(1%) | ||||
| HEPES | Buffering agents to stablize the culture medium | √ | √ | √(10mM) | √(1mM) | ||||
| 1% N2 | Suppresion of cell differentiation and promotion of cell groeth | √ | √ | √ | √ | ||||
| B27 | Suppresion of cell differentiation and promotion of cell groeth | √(1%) | √(1%, minus vitamin A) | √(minus vitamin A) | √(1%) | √(2%, minus vitamin A) | √(2%) | √(2%, minus Vitamin A) | |
| 1× MEM NEAA | Promotion of cell viability | √ | |||||||
| 1× ITS | Insulin: Promotion of energy metabolism; Transferrin: Carrier of Iron and lowering of oxygen radicals; Selenium: Anti-oxidant | √ | |||||||
| 3 μm CHIR99021 | GSK3β inhibitor to activate Wnt pathway | √ | √ | ||||||
| 100 µg mL−1 primocin | Antibiotics to prevent of microorganism contamination | √ | |||||||
| N-Acetylcysteine | Regulate cell proliferation, differentiation and apoptosis | √(1.25mM) | √(1.25mM) | √(1.25mM) | √(1mM) | √(1.25mM) | √(1.25mM) | √(1.25mM) | |
| 10 nM gastrin | Promotion of expention of stomach cells | √ | √ | √ | √ | √ | √ | ||
| EGF | Ligands of receptor protein tyrosine kinase; promotion of expansion of epithelial cells | √(50ng/ml) | √(50ng/ml) | √(50ng/ml) | √(100ng/ml) | √(50ng/ml) | √(50ng/ml) | √(50ng/ml) | |
| FGF-basic | Fibroblast growth factor; Ligands of receptor protein tyrosine kinase | √(1ng/ml) | |||||||
| FGF7 | Fibroblast growth factor; Ligands of receptor protein tyrosine kinase | √(100ng/ml) | √(50ng/ml) | ||||||
| FGF10 | Fibroblast growth factor; Ligands of receptor protein tyrosine kinase | √(100ng/ml) | √(100ng/ml) | √(50ng/ml) | √(100ng/ml) | √(100ng/ml) | √(100ng/ml) | √(100ng/ml) | |
| HGF | Hepatic growth factor | √ (25ng/ml) | √(50ng/ml) | √ (25ng/ml) | √ (25ng/ml) | √ (25ng/ml) | √(50ng/ml) | √(25ng/ml) | |
| RSPO1 conditioned media | Wnt activator | √(10%) | √(15%) | √(15%) | √(10%) | √(10%) | √(10%) | ||
| 10mM Nicotinamide | Sirtuins inhibitor to promote of self-renewal of HCC stem cells and cell energy metabolism | √ | √ | √ | √ | √ | √ | √ | |
| A8301 | TGFβ inhibitor to promote cell expansion and inhibit apoptosis and differentiation | √(5uM) | √(2uM) | √(1uM) | √(3uM) | √(5uM) | √(5μM) | ||
| 10 μM Forskolin | Rho Inhibitor (cAMP activator) | √ | √ | √ | √ | ||||
| 10 μM γ-27632 | Rock inhibitor to inhibit apoptosis of stem cells | √ | √ | √ | √ | √ | |||
| Noggin | BMP-4 and BMP-7 inhibitor to inhibit the differentiation of stem cells | √ (25 ng/ml) | √ (25 ng/ml) | √(10%) | √(5%) | ||||
| 30% Wnt Conditioned medium | Ligand of canonical Wnt/β-catenin pathway to promote cell expansion | √ | √ | √ | √ | ||||
| 20ng/ml TGF-α | Regulators of cell proliferation and differentiation | √ | |||||||
| 10ng/ml Oncostatin M | Inhibitors of proliferation of a number of tumor cell lines | √ | |||||||
| Dexamethasone | Induction of apoptosis of hepatic cancer cells | √(1uM) | √(3uM) | ||||||
BME2, Basement Membrane Extract, Type 2; NEAA, Non-Essential Amino Acids; ITS, Insulin-Transferrin-Selenium; EGF, epidermal growth factor; FGF, fibroblast growth factor; HGF, hepatocyte growth factor; TGFα, transforming growth factor alpha; BMP, bone morphogenetic protein.
√ indicates that the culture medium contains the relevant components.