Abstract
Cultivated meat is an emerging field, aiming to establish the production of animal tissue for human consumption in an in vitro environment, eliminating the need to raise and slaughter animals for their meat. To realise this, the expansion of primary cells in a bioreactor is needed to achieve the high cell numbers required. The aim of this study was to develop a scalable, microcarrier based, intensified bioprocess for the expansion of bovine adipose-derived stem cellsas precursors of fat and muscle tissue. The intensified bioprocess development was carried out initially in spinner flasks of different sizes and then translated to fully controlled litre scale benchtop bioreactors. Bioprocess intensification was achieved by utilising the previously demonstrated bead-to-bead transfer phenomenon and through the combined addition of microcarrier and medium to double the existing surface area and working volume in the bioreactor. Choosing the optimal time point for the additions was critical in enhancing the cell expansion. A significant fold increase of 114.19 ± 1.07 was obtained at the litre scale in the intensified bioprocess compared to the baseline (**p < .005). The quality of the cells was evaluated pre- and post-expansion and the cells were found to maintain their phenotype and differentiation capacity.
Highlights
Over the past couple of years, the concept of cultivated meat has received increased attention due to its undeniable potential as a sustainable food source, as well as its ability to address many of the existing challenges and detrimental effects of livestock meat
It has been estimated that 1011 cells are needed to make 1 kg of meat, including all cell types involved (Post et al, 2020). Such cell numbers are only attainable in bioreactor cultures
Microcarrier culture in bioreactors has a significant advantage over monolayer culture due to their high surface‐area‐ to‐volume ratio
Summary
The step for scalable production is translation to fully controlled bioreactors This is, to our knowledge, the first report of litre scale expansion of bASCs for cultivated meat production. Microcarrier culture in bioreactors has a significant advantage over monolayer culture due to their high surface‐area‐ to‐volume ratio This surface area can be expanded in bioprocesses through the provision of additional microcarriers, maximising cell expansion, while maintaining cell growth in the exponential phase for longer. We investigated the surface area addition approach for process intensification, while translating the bioprocess for expansion of bASCs from spinner flasks to litre scale bench‐top bioreactors, benefiting from full environmental monitoring and control. Such work has not been reported before for bASCs culture and represents a major step forward for this novel use of stem cell culture
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