Research Of MDCK Cell Culture-based Influenza Vaccine Production

Influenza is one of infectious diseases which humanity has noteffectively control. Influenza is a threat to human health worldwide.Although there are antiviral drugs, but drugs may induce drug-resistantstrains. It is not suitable for a wide range to resistant to the influenzavirus. The vaccine is an important method of prevention and control ofinfluenza. In addition to the traditional chick embryo preparation ofinfluenza vaccine, continuous cell lines such as MDCK cells, Vero cells,RER.C6cells have also been used in the preparation of influenzavaccine. MDCK cells is easy to culture, sensitive to a variety of differentsubtypes of influenza virus strains and high yield product of influenzacultured in MDCK cells. It is an ideal cell line for influenza vaccineproduction.This article describes the establishment of three levels of MDCKcells and use this seed bank to carry out research on the growth ofMDCK cells. Different kinds of serum and different concentration ofglucose was adjusted to determine the conditions of MDCK cellsadherent growth in flask. On this basis, I further assessed the growth ofMDCK cells in adherent culture on the microcarrier surface in thebioreactor system. MDCK cells adherent cultured on microcarriers inthe initial inoculation cell density of2×105cells/mL without replacingthe medium. After about72hours incubation, the cell density can reach（13.77±0.69）×105cells/mL×105cells/mL and amplification ratio canreach6.89times. In addition, I optimized the conditions for culturing ofinfluenza virus in MDCK cells. When the cells cultured for72hours,Cell was infected with virus at a MOI of0.01in MEM mediumcontaining2.5μg/mL TPCK-trypsin. The influenza virus HA titers reached9log2HA/50μL after48hours inculation. These results wereimportant to provide technique and theory basis for esteblished atechnology platform of large-scale cultivation of influenza vaccine in thebioreactor microcarrier system.