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Cultured meat is an emerging technology that could address environmental, health, and animal welfare concerns associated with meat production. Development of cultured meat represents an exciting challenge for cell biologists and engineers, but it requires effective, open approaches for knowledge sharing to establish a fertile scientific field alongside a competitive industry. [ABSTRACT FROM AUTHOR]

Metabolic engineering of mammalian cells has to-date focused primarily on biopharmaceutical protein production or the manipulation of native metabolic processes towards therapeutic aims. However, significant potential exists for expanding these techniques to diverse applications by looking across the taxonomic tree to bioactive metabolites not synthesized in animals. Namely, cross-taxa metabolic engineering of mammalian cells could offer value in applications ranging fromfood and nutrition to regenerative medicine and gene therapy. Towards the former, recent advances in meat production through cell culture suggest the potential to produce meat with fine cellular control, where tuning composition through cross-taxa metabolic engineering could enhance nutrition and food-functionality. Here we demonstrate this possibility by engineering primary bovine and immortalized murine muscle cells with prokaryotic enzymes to endogenously produce the antioxidant carotenoids phytoene, lycopene and β-carotene. These phytonutrients offer general nutritive value and protective effects against diseases associated with red and processed meat consumption, and so offer a promising proof-of-concept for nutritional engineering in cultured meat. We demonstrate the phenotypic integrity of engineered cells, the ability to tune carotenoid yields, and the antioxidant functionality of these compounds in vitro towards both nutrition and food-quality objectives. Our results demonstrate the potential for tailoring the nutritional profile of cultured meats. They further lay a foundation for heterologous metabolic engineering of mammalian cells for applications outside of the clinical realm. • Meat production via cell culture can enable tight product composition control. • Phytonutrient production in muscle cells can improve cultured meat nutrition. • The carotenoids phytoene, lycopene, and β-carotene are promising engineering targets. • Carotenoid production was achieved at sufficient levels for antioxidant function. • Tissue-engineered food offers new application for mammalian metabolic engineering. [ABSTRACT FROM AUTHOR]

The development of cost-effective serum-free media is essential for the economic viability of cultured meat. A key challenge facing this goal is the high-cost of recombinant albumin which is necessary in many serum-free media formulations, including a recently developed serum-free medium for bovine satellite cell (BSC) culture termed Beefy-9. Here we alter Beefy-9 by replacing recombinant albumin with rapeseed protein isolate (RPI), a bulk-protein solution obtained from agricultural waste through alkali extraction (pH 12.5), isoelectric protein precipitation (pH 4.5), dissolution of physiologically soluble proteins (pH 7.2), and concentration of proteins through 3 kDa ultrafiltration. This new medium, termed Beefy-R, was then used to culture BSCs over four passages, during which cells grew with an average doubling time of 26.6 h, showing improved growth compared with Beefy-9. In Beefy-R, BSCs maintained cell phenotype and myogenicity. Together, these results offer an effective, low-cost, and sustainable alternative to albumin for serum-free culture of muscle stem cells, thereby addressing a key hurdle facing cultured meat production. [ABSTRACT FROM AUTHOR]

With rising global demand for food proteins and significant environmental impact associated with conventional animal agriculture, it is important to develop sustainable alternatives to supplement existing meat production. Since fat is an important contributor to meat flavor, recapitulating this component in meat alternatives such as plant based and cell cultured meats is important. Here, we discuss the topic of cell cultured or tissue engineered fat, growing adipocytes in vitro that could imbue meat alternatives with the complex flavor and aromas of animal meat. We outline potential paths for the large scale production of in vitro cultured fat, including adipogenic precursors during cell proliferation, methods to adipogenically differentiate cells at scale, as well as strategies for converting differentiated adipocytes into 3D cultured fat tissues. We showcase the maturation of knowledge and technology behind cell sourcing and scaled proliferation, while also highlighting that adipogenic differentiation and 3D adipose tissue formation at scale need further research. We also provide some potential solutions for achieving adipose cell differentiation and tissue formation at scale based on contemporary research and the state of the field. [ABSTRACT FROM AUTHOR]

Scaffolds suitable for use in food products are crucial components for the production of cultured meat. Here, wheat glutenin, an inexpensive and abundant plant-based protein, was used to develop 3D porous scaffolds for cultured meat applications. A physical cross-linking method based on water annealing was developed for the fabrication of porous glutenin sponges and fibrous aligned scaffolds. The pore sizes ranged from 50 to 250 μm, with compressive modulus ranges from 0.5 to 1.9 kPa, depending on the percentage of glutenin (2%–5%) used in the process. The sponges were stable in PBS with refrigeration for at least six months after water annealing. The glutenin scaffolds supported the proliferation and differentiation of C2C12 mouse skeletal myoblasts and bovine satellite cells (BSCs) without the need to add specific cell adhesive proteins or other coatings. The low cost and food safe production process avoided the use of toxic cross-linkers and animal-derived extracellular matrix (ECM) coatings, suggesting that this as approach is a promising system for scaffolds useful in cultivated meat applications. [ABSTRACT FROM AUTHOR]