This paper mainly studies the high-yield characteristics and passage stability of recombinant human infection with highly pathogenic avian influenza H7N9 strain as the main working seed batch in the continuous passage of Vero cells, providing preliminary research for vaccine development.The H7N9 influenza virus Vero cell adaptation group (A/Anhui/1/2013Va) was inoculated into Vero cells at MOI=0.01, and passaged for 15 consecutive generations. Hemagglutination test was used to detect the hemagglutination titer of each generation of virus was and conducted the virus growth curve on Vero cells was drawn; MDCK cells are used for each generation of virus to detect the infectious titer of the virus; type identification of recombinant viruses H7N9 passage 1 and passage 16 was done by single radial immunodiffusion; take the 1st, 5th and 16th generation viruses were taken, and half the amount of infection and lethality of chicken embryos to detect the virulence of the virus during the passage; RNA from H7N9 influenza viruses of the 1st and 16th generation were extracted, reverse-transcribed and sequenced after cloning to compare changes in hemagglutinin(HA) and neuraminidase(NA) genes of the virus during successive passages.The test results showed that the virus titer became more and more stable during successive passages, and the titer remained stable at 384~448; the infectious titer of the virus was maintained at 7 Log10TCID50/mL; virus virulence did not change significantly during the passaging process; HA and NA did not show changes in the encoded amino acids due to gene mutations during continuous passaging. The H7N9 Anhui strain virus has not only a stable yield during the passage of Vero cells, but also its virulence and genes are stable. It can be used as a candidate strain of H7N9 influenza vaccine.