Cascade Oxygen Control Enhances Growth of Nicotiana benthamiana Cell Cultures in Stirred-Tank Bioreactors

Plant cell cultures offer a promising platform for producing valuable biomolecules, yet their use in bioreactors remains under-optimized. Compared to animal or microbial cells, plant cells grow more slowly, limiting productivity. A common bioprocess strategy to improve yields involves controlling dissolved oxygen (DO) levels. However, little research has focused on combining agitation and aeration to regulate oxygen in plant cell cultures within bioreactors. The aim of this study was to evaluate the impact of an oxygen cascade mixing agitation and aeration on plant cell growth in stirred-tank systems. By maintaining 30% DO through this approach, the specific growth rate (µ) increased from 0.082 d^-1 to 0.144 d^-1 on average in <i>Nicotiana benthamiana</i> cultures at the 2 L scale, decreasing batch lengths from 21 to 10 days. These conditions were successfully replicated in a 7 L stainless-steel pilot bioreactor using previous values of geometry, <i>k</i>_L<i>a</i> and <i>N</i>_RE as scale-up criteria, obtaining a µ of 0.161 d^-1. These results demonstrate that plant cell cultures' efficiency can be enhanced by using standard bioprocess parameters. While this work confirms the feasibility of cascade oxygen control for improvements in growth, further studies are needed to evaluate its specific impact on biomolecule production across different systems.