Cultured meat (CM) production has emerged as a breakthrough technology for the global food industry, with the potential to reduce challenges associated with environmental sustainability, global public health, animal welfare, and competition for food between humans and animals. The muscle stem cell lines currently used for CM production cannot be passaged in vitro for extended periods of time. Here, we successfully developed a directional differentiation system of porcine pregastrulation epiblast stem cells (pgEpiSCs) with stable cellular features and achieved serum-free myogenic differentiation of the pgEpiSCs. The pgEpiSCs-derived skeletal muscle progenitor cells and skeletal muscle fibers show typical muscle cell characteristics and display skeletal muscle transcriptional features during myogenic differentiation. Importantly, we further established a three-dimensional differentiation system for shaping cultured tissue by screening plant-based edible scaffolds of non-animal origin, followed by the generation of pgEpiSCs-derived CM. These advances provide a novel technical approach for the development of CM.
