Abstract
Development of serum-free suspension cell culture processes is very important for influenza vaccine production. Previously, we developed a MDCK suspension cell line in a serum-free medium. In the present study, the growth kinetics of suspension MDCK cells and influenza virus production in the serum-free medium were investigated, in comparison with those of adherent MDCK cells in both serum-containing and serum-free medium. It was found that the serum-free medium supported the stable subculture and growth of both adherent and suspension cells. In batch culture, for both cell lines, the growth kinetics in the serum-free medium was comparable with those in the serum-containing medium and a commercialized serum-free medium. In the serum-free medium, peak viable cell density (VCD), haemagglutinin (HA) and median tissue culture infective dose (TCID50) titers of the two cell lines reached 4.51×106 cells/mL, 2.94Log10(HAU/50 μL) and 8.49Log10(virions/mL), and 5.97×106 cells/mL, 3.88Log10(HAU/50 μL), and 10.34Log10(virions/mL), respectively. While virus yield of adherent cells in the serum-free medium was similar to that in the serum-containing medium, suspension culture in the serum-free medium showed a higher virus yield than adherent cells in the serum-containing medium and suspension cells in the commercialized serum-free medium. However, the percentage of infectious viruses was lower for suspension culture in the serum-free medium. These results demonstrate the great potential of this suspension MDCK cell line in serum-free medium for influenza vaccine production and further improvements are warranted.
Highlights
In recent years, animal cell culture technology has gradually replaced the traditional chick embryo production process for influenza vaccine production
The adherent MDCK cells were adapted to suspension culture by the serum reduction and serial passaging approach in another proprietary serum-free medium developed by the authors (MDCK-SFM2) [26, 27], and the resulting suspension MDCK cells were cultured in MDCK-SFM2 or a commercialized serum-free medium Ex-cell MDCK (Sigma-Aldrich)
Stable passaging of MDCK cells under different culture conditions To evaluate the adaptability of MDCK cells, cell growth was investigated in serum-containing adherent, serum-free adherent and serum-free suspension culture modes for 10 passages
Summary
Animal cell culture technology has gradually replaced the traditional chick embryo production process for influenza vaccine production. Most of cells applied for influenza vaccine production are adherent and grown as monolayers. Largescale culture processes mainly rely on cultivating adherent cells on microcarriers in serum-containing medium [1,2,3]. Serum-Free Suspension Culture and Influenza Vaccine Production
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