Continuous culture method and culture medium for primary hepatocellular carcinoma organoids
A technology for culture medium and hepatocytes, which can be used in cell culture active agents, hepatocytes, detection of programmed cell death, etc., can solve the problems of low culture success rate, low success rate, and difficulty in culture
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Embodiment 1
[0043] Embodiment 1, improved culture medium 1
[0044] DMEM / F12 medium, ascorbic acid (10mg / L), L-glutathione (1-50mg / L), Insulin (20mg / L), Sodium selenite (0.05mg / L), R-spondin 1 (500ng / ml), N2(1X), B27(1X), GlutaMAXSupplement(1X), HEPES(1X), Niacinamide(20mM), Normocin TM (1X), N-Acetylcysteine (2mM), A83-01 (10μM), Gastrin I (10nM), EGF (20ng / ml), HGF (50ng / ml), FGF7 (20ng / ml) , FGF10 (200 ng / ml), Y27632 (50 μM).
Embodiment 2
[0045] Embodiment 2, improved culture medium 2.
[0046] DMEM / F12 medium, ascorbic acid (10mg / L), Holo-Transferrin (50mg / L), L-glutathione (20mg / L), ethanolamine hydrochloride (30mg / L), R-spondin 1:( 200ng / ml), N2(1X), B27(2X), GlutaMAXSupplement(1X), HEPES(1X), Nicotinamide(5mM), Normocin TM (1X), N-Acetylcysteine (2mM), A83-01 (5μM), Gastrin I (10nM), EGF (50ng / ml), FGF7 (100ng / ml), FGF10 (100ng / ml) , DMSO (1%), Y27632 (50 μM).
Embodiment 3
[0047] Embodiment 3, improved culture medium 3
[0048] DMEM / F12 medium, Holo-Transferrin (10mg / L), L-glutathione (1-10mg / L), Insulin (10mg / L), R-spondin 1: (100ng / ml), N2 (0.5 X): B27(1X), GlutaMAX Supplement(1X), HEPES(1X), Nicotinamide(10mM), Normocin TM (1X), N-acetylcysteine (1.25 mM), A83-01 (0.5 μM), gastrin I (1 nM), EGF (100 ng / ml), Y27632 (50 μM), CHIR99021 (50 μM).
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