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Abstract

Before considering intracellular metabolites clear ideas on the extracellular conditions of experiments are obligatory. Many interesting questions on understanding metabolism of cells are generated under process conditions and not under defined steady state conditions with defined media. Here, we discuss limits and benefits of the interpretation of extracellular metabolite data for a process of influenza vaccine production in mammalian cells. Therefore, the production process is introduced together with relevant details on medium composition and cultivation conditions. Typical issues of metabolism during cell growth and viral infection are presented. Data on basic metabolites (glucose, lactate, glutamine, ammonia) are shown together with amino acid profiles measured from the cell culture medium [1]. Comparing different media (serum-containing, glutamine-free, serum-free [2-4]) as well as different cultivation methods (roller bottles, stirred tank bioreactor, wave bioreactor) it is demonstrated, what can be learned from extracellular metabolites and what questions remain. [1] Genzel Y.; König S.; Reichl U.; "Amino acid analysis in mammalian cell culture media containing serum and high glucose concentrations by anion exchange chromatography and integrated pulsed amperometric detection (IPAD)." , Anal. Biochem. 2004, 335, 119-125. [2] Genzel Y.; Behrendt I.; König S.; Sann H.; Reichl U.; "Metabolism of MDCK cells during cell growth and influence virus production in large-scale microcarrier culture" , Vaccine 2004, 22(17-18), 2202-2208. [3] Genzel Y.; Ritter JB.; König S.; Alt R.; Reichl U.; "Substitution of glutamine by pyruvate to reduce ammonia formation and growth inhibition of mammalian cells." , Biotechnol. Progr. 2005, 21 (1), 58-69. [4] Genzel Y.; Fischer M.; Reichl U.; "Serum-free influenza virus production avoiding washing steps and medium exchange in large-scale microcarrier culture.", Vaccine 2006, 24(16), 3261-3272.