ObjectiveTo provide theoretical and technological support for further study of liver metabolism and disease by comparing the advantages and disadvantages of various artificial liver models (biological). MethodsLiteratures were searched and compared to summarize the requirement for liver donor, isolation, and culture of hepatocyte. ResultsIn the separation method of hepatocyte, mechanical separation method had no requirement for liver donor, and was easy to acquire hepatocyte, while the acquired hepatocyte would be destructed severely, and the survival rate was low. On the other hand, the restriction of the digestion of the hepatocytes to the liver cell samples was unlimited, while the key of the enzyme digestion method was to regulate the balance between enzyme concentration and digestion time, which was limited to function researches of hepatocyte, and research about the responds of hepatocyte against outside, and other few researches. Perfusion digestion method had been widely applied for animal test. The Ca2+, collagenase and perfusion rate, pH value, buffer, and intubation method all play vital roles. During the cultivation, we needed to choose different methods according to several experiments, and add different additives in the appropriate medium. Different biological reactors had different advantages, disadvantages, and applicable conditions. ConclusionsThe donor selection is based on various experimental purposes to harvest hepatocytes from different sources. Whether on the separation process or on the cultivation process, according to the specific circumstances, such as the concentration, perfusion time, and the choice of different kinds of culture medium, we can choose different kinds of bioreactors, but all kinds of methods are still remained with multiple insufficiencies, which require more researchers to improve.