Serum-Free Suspension Culture of MDCK Cells for Production of Influenza H1N1 Vaccines

• Published in: PloS one 2015-11, Vol.10 (11), p.e0141686-e0141686
• Main Authors: Huang, Ding, Peng, Wen-Juan, Ye, Qian, Liu, Xu-Ping, Zhao, Liang, Fan, Li, Xia-Hou, Kang, Jia, Han-Jing, Luo, Jian, Zhou, Lin-Ting, Li, Bei-Bei, Wang, Shi-Lei, Xu, Wen-Ting, Chen, Ze, Tan, Wen-Song
• Format: Article
• Language: English
• Subjects: Adherent cells; Animals; Apoptosis; Batch Cell Culture Techniques - methods; Batch culture; Biological products; Biotechnology; Cell Count - methods; Cell culture; Cell density; Cell growth; Cell Line; Cell lines; Commercialization; Culture Media, Serum-Free - pharmacology; Dogs; Engineering; Growth kinetics; Hemagglutinins; Hemagglutinins - immunology; Influenza; Influenza A Virus, H1N1 Subtype - immunology; Influenza Vaccines - biosynthesis; Influenza Vaccines - immunology; Kinetics; Laboratories; Madin Darby Canine Kidney Cells; Orthomyxoviridae; Orthomyxoviridae Infections - immunology; Orthomyxoviridae Infections - prevention & control; Productivity; Reaction kinetics; Science; Serum-free medium; Subculture; Suspension culture; Swine flu; Tissue culture; Vaccines; Virions; Virus Cultivation - methods; Viruses
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0141686

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.