Proteome constrained metabolic modeling of Sus scrofa muscle stem cells for cultured meat production

Abstract Cultured meat has recently emerged as a sustainable alternative to the traditional livestock farming and gained attention as a promising future protein source. Herein, the Sus scrofa muscle stem cell is a commonly used cell source in the cell proliferation step of cultured meat production. However, a major bottleneck of large-scale cultivation is the inhibition by secreted and accumulated lactate and ammonium in the process of S. scrofa cell proliferation. To simulate the growth and metabolism of S. scrofa muscle stem cells under different lactate and ammonium concentrations, this study constructed the first proteome constrained metabolic model for the core metabolism of S. scrofa muscle stem cells, pcPigGEM2025. The relationship of lactate and ammonium levels with cellular metabolism was derived from growth and metabolomics data of two culture conditions with low and high initial ammonium concentrations, and then incorporated into metabolic flux simulation. Metabolic flux simulations for experimental conditions, along with perturbation simulations considering stressed non-growth associated maintenance and oxygen supply, demonstrated that pcPigGEM2025 could effectively characterize the response of the S. scrofa muscle stem cell’s growth and metabolism to varying environmental conditions, shedding light on model-aided control and optimization of the cultured meat production process. Highlights The first proteome constrained metabolic model was built for S. scrofa myoblasts. This model effectively simulated myoblast metabolism under lactate and NH4+ stress. Perturbation simulations showed that this model could also account for other stress. This model enables in-silico control and optimization of cultured meat production. Competing Interest Statement Prof. Hua Ye is the co-founder of Ivy Farm Technologies (Oxford, UK). Footnotes Abbreviations - CHO - Chinese hamster ovary - DCW - dry cellular weight - DW - dry weight - GAM - growth associated maintenance - GEM - genome-scale metabolic model - GPR - gene-protein-reaction rule - FBA - flux balance analysis - NGAM - non-growth associated maintenance - NH4+ - ammonium - OUR - oxygen uptake rate - TCA - tricarboxylic acid