Scale‐up of an intensified bioprocess for the expansion of bovine adipose‐derived stem cells (bASCs) in stirred tank bioreactors

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 cells as 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.

Scaffolding Biomaterials for 3D Cultivated Meat: Prospects and Challenges

“…Although substantial progress has been achieved in recent years by both for‐profit companies and academic groups, CM remains a relatively new area of investigation. As might be expected for a nascent field that draws heavily on knowledge gained from previous work aimed at a distinct set of problems, review, and perspective papers about CM (at least 50 by our estimate) [ 10–14,16,18,23,28,37,44,64–109 ] currently outnumber original research papers where CM production is the primary intended application (at least 24) [ 15,59,110–131 ] or CM‐related patents (at least 24). [ 132–155 ] These estimates do not include LCA or TEA studies, consumer acceptance studies, or purely computational analyses.…”

Needle to needle robot‐assisted manufacture of cell therapy products

“…Cell quality postexpansion was assessed by multilineage differentiation and flow cytometry for cell surface marker expression. The multilineage differentiation capacity of the hMSC cultures was assessed by their differentiation to the classical MSC lineages, that is, chondrogenic, osteogenic, and adipogenic using established protocols 36 . All reagents were supplied by Sigma Aldrich unless stated otherwise.…”

Bioengineering Outlook on Cultivated Meat Production

“…Hähnel and co-workers have evaluated several commercially available disposable STRs for mammalian cell cultivation [ 165 ]. A single-use, disposable STR vessel (Mobius CellReady 3 L, MilliporeSigma, Burlington, MA, USA) was recently applied for the expansion of bovine adipose-derived stem cells (bASCs) as precursors of fat and muscle tissue [ 167 ]. This study has given new perspectives on CM production by increasing the scale of a microcarrier-based (SoloHill Plastic microcarriers, Pall Corporation, New York, NY, USA) expansion bioprocess from 100 mL-spinner flask across 500 mL-spinner flask to 3 L-STR with a disposable vessel.…”

“…This study has given new perspectives on CM production by increasing the scale of a microcarrier-based (SoloHill Plastic microcarriers, Pall Corporation, New York, NY, USA) expansion bioprocess from 100 mL-spinner flask across 500 mL-spinner flask to 3 L-STR with a disposable vessel. Furthermore, the process intensification approach based on “surface + volume addition”, i.e., timely-defined addition of fresh MCs to support bead-to-bead transfer and medium exchange, has resulted in a significant fold increase of 114.19 ± 1.07 when it comes to bASC cell number [ 167 ].…”