Optimization of slaughter livestock serum as an alternative to fetal bovine serum in cultured meat application

Fetal bovine serum (FBS) is widely used for cultured meat production, but its fetal origin raises ethical concerns and contributes to high cost and batch variability, driving the demand for practical alternatives. This study evaluated livestock sera from bovine, porcine, and chicken sources through biochemical profiling, proliferation and differentiation assays, and long-term serial passaging of bovine satellite cells. All sera met sterility requirements, although some livestock sera showed deviations in physicochemical parameters. Bovine and porcine sera, unlike chicken serum, consistently supported robust proliferation, myogenic differentiation, and long-term expansion. Importantly, the final FBS substitutes were optimized through supplementation with lipid-rich albumin or insulin–transferrin–selenium (25 µg/mL), which significantly enhanced cellular performance. Cost analysis revealed that replacing FBS with the optimized livestock-derived serum formulations reduced total medium cost by 62.6%. Collectively, optimized bovine- and porcine-based FBS substitutes represent effective, scalable, and ethically aligned alternatives for sustainable cultured meat bioprocessing. Key points • Bovine and porcine sera fully replaced FBS for bovine satellite cells • Optimized FBS substitutes supported stable proliferation during serial passaging • Replacing FBS reduced total culture medium cost by 62.6%