title: Serum-Free Suspension Culture of MDCK Cells for Production of Influenza H1N1 Vaccines.
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.
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
authors.forename: Ding
authors.initials: D
authors.lastname: Huang
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
authors.forename: Wen-Juan
authors.initials: WJ
authors.lastname: Peng
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Ye
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
authors.forename: Xu-Ping
authors.initials: XP
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
authors.forename: Liang
authors.initials: L
authors.lastname: Zhao
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
authors.forename: Li
authors.initials: L
authors.lastname: Fan
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
authors.forename: Kang
authors.initials: K
authors.lastname: Xia-Hou
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
authors.forename: Han-Jing
authors.initials: HJ
authors.lastname: Jia
authors.affiliation: Shanghai Institute of Biological Products Co., Ltd., Shanghai 200052, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Luo
authors.affiliation: Shanghai Institute of Biological Products Co., Ltd., Shanghai 200052, China.
authors.forename: Lin-Ting
authors.initials: LT
authors.lastname: Zhou
authors.affiliation: Shanghai Institute of Biological Products Co., Ltd., Shanghai 200052, China.
authors.forename: Bei-Bei
authors.initials: BB
authors.lastname: Li
authors.affiliation: Shanghai Institute of Biological Products Co., Ltd., Shanghai 200052, China.
authors.forename: Shi-Lei
authors.initials: SL
authors.lastname: Wang
authors.affiliation: Shanghai Institute of Biological Products Co., Ltd., Shanghai 200052, China.
authors.forename: Wen-Ting
authors.initials: WT
authors.lastname: Xu
authors.affiliation: Shanghai Institute of Biological Products Co., Ltd., Shanghai 200052, China.
authors.forename: Ze
authors.initials: Z
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
authors.forename: Wen-Song
authors.initials: WS
authors.lastname: Tan
mesh_terms: D000818:Animals; D061249:Batch Cell Culture Techniques / Q000379:methods; D002452:Cell Count / Q000379:methods; D002460:Cell Line; D016895:Culture Media, Serum-Free / Q000494:pharmacology*; D004285:Dogs; D006388:Hemagglutinins / Q000276:immunology; D053118:Influenza A Virus, H1N1 Subtype / Q000276:immunology*; D007252:Influenza Vaccines / Q000096:biosynthesis* / Q000276:immunology*; D061985:Madin Darby Canine Kidney Cells; D009976:Orthomyxoviridae Infections / Q000276:immunology* / Q000517:prevention & control*; D014776:Virus Cultivation / Q000379:methods
chemical_list: D016895:Culture Media, Serum-Free; D006388:Hemagglutinins; D007252:Influenza Vaccines