Vaccine is the most effective way to prevent and control infectious diseases. Approximately 80% of infectious diseases are viral diseases. The traditional viral vaccines are prepared by virus cultivation, antigen harvest, antigen concentration, purification and filtration process. Since the growth and reproduction of viruses must take place in the cells, the quality of the cell substrates directly affects the quality of the vaccine products. This article reviews the current status of various cell substrates and discuss their advantages and disadvantages, aiming to provide a reference for choosing the appropriate cell substrate for human vaccine production.