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.

Development of high-growth influenza H7N9 prepandemic candidate vaccine viruses in suspension MDCK cells

“…1B). Several suspension MDCK cell lines have also been developed to facilitate cell-based manufacturing processes [31][32][33]. Similar to these studies, the productivity of influenza viruses in sMDCK cells was higher than that in aMDCK cells (Fig.…”

Cell Bank Origin of MDCK Parental Cells Shapes Adaptation to Serum-Free Suspension Culture and Canine Adenoviral Vector Production

“…Medium formulation used in each case is indicated in each box on the bottom-right corner. The cells used in this work for transcriptome analysis are indicated with ‘*’ [ 9 , 19 , 32 , 46 , 47 , 48 , 49 , 50 , 51 , 52 , 53 ].

Figure A5 Gating parameters on the monitoring of infection progression by flow cytometry analysis.

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Beneficial Effect of Indigo Naturalis on Acute Lung Injury Induced by Influenza a Virus

“…In vitro antiviral evaluation was conducted as described with minor modi cation [19]. Madin-Darby canine kidney (MDCK) cells were cultured in DMEM supplemented with 10% fetal bovine serum (Hyclone, CA, USA), streptomycin and penicillin.…”