Culture method for long-term maintenance and proliferation subculture of human hepatocytes

A method for culturing hepatocytes, involving maintenance, proliferation, and passaging thereof, wherein the method comprising:

• culturing hepatocytes by a culture medium,
• wherein said culture medium comprises:
• a hepatocyte basic medium;
• a GSK-3 beta inhibitor at a concentration of 0.5-10 μM, said GSK-3 beta inhibitor is CHIR99021;
• a TGF beta inhibitor at a concentration of 0.5-10 μM, said TGF beta inhibitor is SB 431542 or A83-01;
• N-acetyl-cysteine at a concentration of 0.25-25 mM;
• and
• Oncostatin M at a concentration of 1-100 ng/mL,
• wherein there is no exogenous gene being introduced into the hepatocytes, and the genetic background of the hepatocytes is not changed.