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Among future food problems, the demand for meat is expected to increase rapidly, but the production efficiency of meat, which is a protein source, is very low compared to other foods. To address this problem, research on the development and production of cultured meat as an alternative meat source using muscle stem cells in vitro has recently been undertaken. Many studies have been conducted on myosatellite cells for medical purposes, but studies on alternative meat production are rare. In vitro cell culture mimics the in vivo environment for cell growth. The satellite cell niche is closer to hypoxic (2% O₂) than normoxic (20% O₂) conditions. The aim of this study was to investigate the efficient oxygen conditions of myosatellite cell cultures for the production of cultured meat. The bovine satellite cell counts and mRNA (Pax7, Myf5 and HIF1α) levels were higher in hypoxia than normoxia (p < 0.05). Through Hoechst-positive nuclei counts, and expression of Pax7, MyoD and myosin protein by immunofluorescence, it was confirmed that muscle cells performed normal proliferation and differentiation. Myoblast fusion was higher under hypoxic conditions (p < 0.05), and the myotube diameters were also thicker (p < 0.05). In the myotube, the number of cells was high in hypoxia, and the expression of the total protein amounts, differentiation marker mRNA (myogenin, myosin and TOM20), and protein markers (myosin and TOM20) was also high. The study results demonstrated that the proliferation and differentiation of bovine myosatellite cells were promoted more highly under hypoxic conditions than under normoxic conditions. Therefore, hypoxic cultures that promote the proliferation and differentiation of bovine myosatellite cells may be an important factor in the development of cultured meat. Full article

Skeletal muscle-tissue engineering can be applied to produce cell-based meat for human consumption, but growth parameters need to be optimized for efficient production and similarity to traditional meat. The addition of heme proteins to plant-based meat alternatives was recently shown to increase meat-like flavor and natural color. To evaluate whether heme proteins also have a positive effect on cell-based meat production, bovine muscle satellite cells (BSCs) were grown in the presence of hemoglobin (Hb) or myoglobin (Mb) for up to nine days in a fibrin hydrogel along 3D-printed anchor-point constructs to generate bioartificial muscles (BAMs). The influence of heme proteins on cell proliferation, tissue development, and tissue color was analyzed. We found that the proliferation and metabolic activity of BSCs was significantly increased when Mb was added, while Hb had no, or a slightly negative, effect. Hb and, in particular, Mb application led to a very similar color of BAMs compared to cooked beef, which was not noticeable in groups without added heme proteins. Taken together, these results indicate a potential benefit of adding Mb to cell culture media for increased proliferation and adding Mb or Hb for the coloration of cell-based meat. Full article