Serum-free media development and validation for cultivation of C2C12 immortalised murine myosatellite cell line for cultivated meat

Abstract: The development of cost-effective, serum-free media is critical for scalable cultivated meat production. This study used Design of Experiments (DoE) and high-throughput screening to develop "MMM1", an animal-free, serum-free medium for the C2C12 murine myosatellite cell line. Low cost, food-grade inputs such as methylcellulose and spirulina extract proved significant growth improvements, but this was reversed by incorporating food-grade methylcellulose and Spirulina extract. The optimised MMM1 formulation achieved cumulative population doublings comparable to 10% (v/v) foetal bovine serum over four passages. Furthermore, MMM1 supported scalable cell expansion on dextran-based microcarriers (Cytodex-3) in spinner flasks, matching growth rates of serum-based controls. Finally, transitioning to a food-grade DMEM/F12 basal medium maintained cell proliferation equivalent to pharmaceutical-grade media, offering a viable strategy to substantially reduce biomanufacturing costs.

submitted to 'Food Chemistry'