The principle of rocking motion was introduced to the field of cell culture bioreactors in the late 90s of the last century. Since then, rocking bioreactor systems have been used successfully for small- and medium-sized applications with various cell types in the biopharmaceutical industry. The latest evolution is the ReadyToProcess WAVE 25 bioreactor system from GE Healthcare. With this system, the rocking speed, angle, and—which is new—the sinusoidal profile can be adjusted. The system can be equipped with bags in the sizes of 2 L, 10 L, 20 L, and 50 L with 1 L, 5 L, 10 L, and 25 L maximum working volumes, respectively. For the design and scale-up of biological processes, the main engineering parameters of a bioreactor system are essential. Therefore, the objective of this study was to determine mixing time, oxygen mass transfer rate, and power input; and subsequently demonstrate the validity of determining these parameters in plant suspension cell cultivations. The mixing time was measured with the decolourization method and the specific oxygen mass transfer coefficient was determined by the gassing out method. Measuring the specific power input of a rocking device cannot be conducted. Hence, computational fluid dynamics simulations were used to determine these data. With this approach, the fluid flow pattern, including shear and normal forces and the Kolmogorov microscale of turbulence, could be calculated. The mixing time was in a range suitable for cell cultures and the oxygen mass transfer coefficient was found to be up to 12 h-1. The specific power input ranged between 50 and 500 W m-3. The ability to change the sinusoidal profile of the rocking motion influences these parameters and will be discussed in this presentation. Based on these findings, mass propagations of fast-growing Nicotiana tabacum BY-2 suspension cells were realized in triplicate runs, showing a reproducible cell growth. A maximum cell fresh weight of 350 g L-1 could be achieved, which is comparable to data found in the literature. Thus, the new ReadyToProcess WAVE 25 bioreactor system, equipped with a 20 L culture bag, is characterized with respect to the main important engineering parameters. The successful cultivation of plant cells demonstrates the system’s suitability for this specific type of applications.

Engineering characterization and cultivation of fast growing plant suspension cells using the ReadyToProcess WAVE™ 25 bioreactor system

253rd National Meeting (2017)

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