Screening Of BHK₂₁ Cell Suspension Cultures And Preparation Of O-type Of FMD With High Titer

The growth of cells requires a medium to provide nutrients.In order to achieve large-scale and industrialization,to achieve precise control of the production process,reduce production costs,facilitate downstream work and improve the quality of biological products and safety,we need to continuously improve cell culture technology,screening appropriate BHK₂₁ cell culture medium.In this study,we selected suitable medium to increase the number and quality of cells by cell number,cell viability,cell apoptosis and cell availability,and further screened the high-titer,stable,poison.The cells were inoculated into A,B,C three kinds of culture medium to suspension culture.The number of cells,apoptosis and cell viability were used to screen the suitable medium for the growth of the cells.The results show:The proportion of total cell apoptosis in culture medium A was 3.5%⁸.2%,which was lower than that of culture medium C,which was much lower than that of culture medium B at32 h,36 h and 48 h after suspension culture.This means that apoptosis has been at a low level and the growth state remained stable.The cells were cultured in suspension medium for 10 h,34 h and 48 h,and the cell availability in culture medium A was 97%⁹8%,higher than that of medium C and B.Therefore,the cells are more suitable for growth and metabolism in culture A.O/Mya98/2010 of FMD were domesticated,then the toxicity of each generation and 146S were determined,and the best generation was screened out.BF9 generation of adherent bottle virus can be used as the production of seed virus for suspension of domestication.BF11 generation of suspended virus can be done as a production of toxic amplification suspension.BF11 generation of suspended virus which was the first time passed was selected for amplification and culture.The suspension virus was cultivated in a bioreactor of 50L.And then it was detected that TCID₅₀average was 10^-7.5/0.1ml,and 146S average was 3.81?g/mL.It was compared with the previous significantly improved which TCID₅₀ was 10^-7.0/0.1ml and 146S was 1.2¹.5?g/mL.So it can achieve the standard of production antigen.