Abstract
Plant cell suspension cultures are widely used for the production of recombinant proteins and secondary metabolites. One of the most important steps during process development is the optimization of yields by testing different cultivation parameters, including the components of the growth medium. However, we have shown that the biomass yield of a cell suspension culture derived from the pear cultivar Pyrus communis cv. Champagner Bratbirne can be significantly improved solely by varying the temperature, inoculum density, illumination, and incubation time. In contrast to medium optimization, these simple physical factors are easily controlled and varied, thereby reducing the effort required. Using an experimental design approach, we improved the biomass yield from 146 g fresh weight (FW)/L to 407 g FW/L in only 5 weeks, simultaneously reducing the costs of goods sold per kg biomass from €125 to €45. Our simple approach therefore offers a rapid, efficient and economical process for the optimization of plant cell suspension cultures.
Highlights
Number of relevant factors that must be tested in combination is much lower than typically required for medium optimization, reducing the complexity of the experimental design and the effort required for screening
We investigated whether any of the -controllable and variable factors light, temperature, incubation time and inoculum density had a significant impact on the accumulation of biomass and whether varying these factors could achieve biomass accumulation similar to BY-2 cells
To assess these four factors, we set up a IV-optimal response surface method (RSM) comprising 35 runs using Design Expert v.8.04 (Supplement 1)
Summary
Number of relevant factors that must be tested in combination is much lower than typically required for medium optimization, reducing the complexity of the experimental design and the effort required for screening. We analyzed the impact of four of the most controlled physical factors (light, temperature, incubation time and inoculum density) on the biomass yield of a pear suspension cell culture by modelling the design space using a DOE approach. This model enabled us to improve the biomass yield 3-fold, and reduced the cost of goods sold (COGS) by the same order, without any attempt to optimize the culture medium. This approach provides a simple and efficient strategy to improve the performance of plant cell suspension cultures and it is likely that the same method could be applied in other cell-based production platforms to increase productivity and reduce the production costs
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