Bioprocess development for scalable production of cultivated meat.

Mariana P Hanga,Panagiota Moutsatsou,Alvin Nienow,Fritz A De La Raga,Junaid Ali,Ivan Wall,Christopher J Hewitt

doi:10.1002/bit.27469

Mariana P Hanga, Panagiota Moutsatsou + Show 5 more

Open Access

https://doi.org/10.1002/bit.27469

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Abstract

Traditional farm-based products based on livestock are one of the main contributors to greenhouse gas emissions. Cultivated meat is an alternative that mimics animal meat, being produced in a bioreactor under controlled conditions rather than through the slaughtering of animals. The first step in the production of cultivated meat is the generation of sufficient reserves of starting cells. In this study, bovine adipose-derived stem cells (bASCs) were used as starting cells due to their ability to differentiate towards both fat and muscle, two cell types found in meat. A bioprocess for the expansion of these cells on microcarriers in spinner flasks was developed. Different cell seeding densities (1,500, 3,000, and 6,000 cells/cm2 ) and feeding strategies (80%, 65%, 50%, and combined 80%/50% medium exchanges) were investigated. Cell characterization was assessed pre- and postbioprocessing to ensure that bioprocessing did not negatively affect bASC quality. The best growth was obtained with the lowest cell seeding density (1,500 cells/cm2 ) with an 80% medium exchange performed (p < .0001) which yielded a 28-fold expansion. The ability to differentiate towards adipogenic, osteogenic, and chondrogenic lineages was retained postbioprocessing and no significant difference (p > .5) was found in clonogenicity pre- or postbioprocessing in any of the feeding regimes tested.

Highlights

There is an increased need for sustainable, protein rich food sources to support the rapidly growing population (Arshad et al, 2017)
The cells were cultured for 10 consecutive passages. bovine adipose-derived stem cells (bASCs) morphology was monitored throughout the entire duration of the continued culture and it was found to be fibroblastlike and similar to that reported for human mesenchymal stem cells (Heathman et al, 2016) (Figure 1A)
In comparison to human mesenchymal stem cells (hMSCs), bASCs were found to be smaller with sizes ranging from 13μm at the earlier passages to 16μm at the later passages (Figure 1C)

Summary

Introduction

There is an increased need for sustainable, protein rich food sources to support the rapidly growing population (Arshad et al, 2017). With animal agriculture currently occupying 70% of arable land, generating 14.5% of anthropogenic greenhouse emissions (Grossi et al, 2019) and consuming 27% of fresh water resources just for livestock feed production (Gerbens-Leenes et al, 2013), it becomes evident that conventional animal agriculture and meat production methods cannot sustain such growth in meat demand Alternative food technologies such as cultivated meat might provide a solution to this growing problem (Salonen & Helne, 2012; Fan et al, 2019; Stephens et al, 2018), as initial projections show that it will require 45% less energy, 99% less land and will emit 78-96% less greenhouse gas emissions (Tuomisto et al, 2011; Stephens et al, 2018). Cultivated meat can undoubtedly have a positive impact on animal welfare and can offer a potentially healthier and safer option for consumers as the production process can be closely controlled and possibly tuned to produce meat that is free from antibiotics, free from zoonotic bacteria and viruses and with a specific, desired nutritional profile (e.g. enriched in omega fatty acids, reduced cholesterol)

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