The invention provides a proliferation culture medium and differentiation culture medium for hepatocyte culture and liver organ preparation. The proliferation culture medium and the differentiation medium both take a culture medium for growth of mammalian cells as a basic culture medium, and an agent for supplementing L-glutamine, a pH value modifier for maintaining the pH values of the culture media stable, a primary cell culture antibiotic, a serum substitute, N-acetylcysteine, arbitrary nicotinamide and any one or more of a BMP inhibitor, a Wnt agonist, a growth factor, a Rock signaling pathway inhibitor, a P38 signal path inhibitor, a Notch signal path inhibitor, dexamethasone, BMP7 and a cAMP activator are added into the culture media. By using the culture media for culturing liver cells, functional liver organs can be obtained.