Improvement of a Laboratory Seed-Train and Characterization of Quail Suspension Cell Line CCX.E10 for Cultivated Meat Production

Abstract

Livestock production forms a threat to the environment and human health, while meat consumption increases due to global population and income growth. Commercial scale cultivated meat production provides a credible alternative due to its resemblance to the taste, texture and looks of conventional meat. In order to commercialize cultivated meat, major technological improvements are needed to scale-up its production from laboratory to commercial scale. This requires an animal cell line that can provide a food-grade product. Quail suspension cell line CCX.E10 is not genetically modified, is spontaneously immortalized and cultivated in animal component free medium. To research the growth kinetics of the cell line for commercial scale production investigation at a laboratory scale is required. Through this research a seed-train protocol to scale-up CCX.E10 cell bank to a scale-down model, namely a DASbox mini bioreactor system, was improved. Additionally, the growth kinetics of CCX.E10 were characterized during batch cultivation, while utilizing this system. A Design-of-Experiments approach was developed to understand what variables have the largest impact on growth kinetics of CCX.E10 to improve operational conditions. Input variables of the approach include the aeration rate, agitation rate and dissolved oxygen concentration of the bioreactor. This research found an exponential specific growth rate of 0.016 h-1 and viable cell density of 2.1x10^6 cells/mL for batch cultivation of cell line CCX.E10 in a DASbox bioreactor. A cell-specific lactate production rate 30% higher than the cell-specific glucose consumption rate was observed during the lag phase of cell cultivation. The growth kinetics derived during cell proliferation suggest that cell line CCX.E10 exhibits the Warburg effect. The findings indicate that the operational conditions for cultivated meat production based on CCX.E10 require further improvement to design a commercial scale bioprocess.