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Kultivierung von MDCK-Zellen und Influenza-A-Virusproduktion im Wave-Bioreaktor


Olmer,  R.
Bioprocess Engineering, Max Planck Institute for Dynamics of Complex Technical Systems, Max Planck Society;
Universität Bielefeld, Lehrstuhl für Zellkulturtechnik;

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Olmer, R. (2004). Kultivierung von MDCK-Zellen und Influenza-A-Virusproduktion im Wave-Bioreaktor. Diploma Thesis, Universität, Bielefeld.

Cite as: https://hdl.handle.net/11858/00-001M-0000-0013-9ED0-D
The production of vaccines in mammalian cell culture processes becomes more and more important. Most of these processes use serumcontaining media for a better cell growth and productivity. However the use of serum is expensive, the lot-to-lot variations, the risk of prion or microorganism contamination as well as high loads of antibodies and complement has lead to a demand of serum free processes. In parallel new cultivation methods have come up, that allow especially for small scale productions and processes from gene therapy the use of presterilized cultivation bags instead of complicated validation procedures for sterilization of cultivation vessels. Here is the adaption of an influenza vaccine production process, using MDCK cells on microcarriers as host system, to the wave-bioreactor presented. Furthermore the comparison of this cultivation in serumcontaining and serumfree medium is shown. Therefore a 5 L standard process for the production of influenza virus A with MDCK cells as host cell line in serumcontaining medium (GMEM) was transferred to the wave-bioreactor. The standard process reaches cell yields of 1 x 106 cells/mL and virus yields of about 2,4 log HA units/100 μL. The cultivations of MDCK in the wave-bioreactor with serumcontaining medium could reach 2 x 106 cells/mL and virus titer of 2,9 log HA units/100 μL. The cultivation of the MDCK-cells on Cytodex 1 microcarriers in Ex-cellTM was not possible without changing some cultivation parameters. Best results gave the cultivation with Ex-cellTM medium enriched with 100 mg Ca2+ and aeration with air + 2 % CO2. In this case cell yields of 1 x 106 cells/mL and virus yields of 2,6 log HA units/100 μL were achieved.