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Abstract:
In cell culture technology high ammonia concentrations can limit cell growth and product formation, especially when focussing on higher cell densities. This high ammonia concentration normally results from glutamine consumption and decomposition. To overcome such problems we have tested several options and developed a glutamine-free medium for cell lines used in industrial applications.
Preliminary investigations were carried out in static cultures of BHK 21, CHO and MDCK cells.
Detailed characterization of MDCK cell metabolism was done for a vaccine production process of influenza virus (equine influenza Newmarket 1/93 H3N8) in MDCK cells using this newly developed glutamine-free medium: The results for cell growth and virus replication in a large-scale controlled microcarrier culture (5L bioreactor) are compared to the production in glutamine containing (2 mM) GMEM medium. Amino acid uptake and release will be compared with other on- & off-line data, to evaluate the importance of some amino acids in different phases of cell metabolism and virus replication with respect to lactate and ammonia production.
