Production of influenza virus in serum-free mammalian cell cultures.
Human influenza viruses are routinely isolated and grown in a variety of mammalian cell substrates. However, influenza viruses for use as inactivated vaccine are still produced in embryonated eggs. Using a perfusion culture-based bioreactor process using serum-free medium, both human and equine influenza viruses of different types and subtypes could be produced to high titres. Classical DEAE-dextran microcarriers were found to be more suitable than polyester sponge carriers for virus production. In addition, MDCK cells grown in serum-free medium were further validated as the most suitable cell substrate compared to Vero and BHK-21 C13 cells for large scale virus production of influenza virus. Finally, to minimize potential contamination by adventitious agents, it was demonstrated that a new serum-free medium in which all animal-derived products are replaced by a plant extract, efficiently supports the growth of MDCK cells as well as the production of influenza virus in the presence of trypsin when using the perfusion bioreactor process.
Merten
O W
OW
Laboratoire de Technologie Cellulaire, Institut Pasteur, Paris, France.
Manuguerra
J C
JC
Hannoun
C
C
van der Werf
S
S
D000818:Animals; D002364:Caseins; D018929:Cell Culture Techniques / Q000379:methods; D002455:Cell Division; D002460:Cell Line / Q000821:virology*; D002522:Chlorocebus aethiops; D006224:Cricetinae; D016895:Culture Media, Serum-Free; D004285:Dogs; D006736:Horses; D006801:Humans; D009980:Influenza A virus / Q000254:growth & development*; D008322:Mammals; D010446:Peptide Fragments; D010936:Plant Extracts; D014709:Vero Cells; D014776:Virus Cultivation
D002364:Caseins; D016895:Culture Media, Serum-Free; D010446:Peptide Fragments; D010936:Plant Extracts; C018135:proteose-peptone