非表示:
キーワード:
-
要旨:
The switch from serum containing media to serum-free media in mammalian cell culture has become a major issue in the last years. Especially for virus vaccine production processes such as influenza, where the infection phase has to be serum-free, even when cultivating the cells in serum containing media, a complete serum-free process has many advantages. From a process engineering point of view the fact that any washing steps as well as addition of fresh media before infection could be omitted, would be a major advantage. Questions that come up are: Do we see any limiting substrates or inhibiting metabolite concentrations during cell growth and virus replication phase? Do the cells attach and grow on microcarriers as good as in serum containing medium? Are commercially available serum-free media, which are only partly optimized for cultivation on microcarriers and often not ideal for achieving maximum virus yields a good candidate for such processes?
Here, we present the successful adaptation of an influenza vaccine production process from serum containing GMEM medium to serum-free Ex-CellTM MDCK medium for microcarrier systems (Cytodex 1). Cultivations in roller bottles, 2L-Wave bioreactor and 5L-stirred tank bioreactor are compared. Online data (pO2, pCO2) as well as offline data such as metabolite profiles for glucose, lactate, glutamine, glutamate and ammonia, cell numbers and virus titers are shown. Advantages and disadvantages as well as changes in metabolite profiles are discussed for serum-free against serum containing medium looking also at different cultivation methods.
