Regulation of myogenic differentiation of myoblasts by groove structure for the design of cell culture meat scaffolds

As a meat substitute, cell cultured meat aims to simulate the composition of real meat to provide nutrient substance. The myotube formation efficiency of myoblasts in vitro is the key for simulating the structure and composition of muscle tissue. Therefore, this study is aimed to explore the most effective groove widths for myoblasts fusion and related molecular mechanism. The results of immunofluorescence showed that the grooves with a width range of 50 μm–1000 μm were beneficial to cell arrangement compared to flat structure. At the same time, the expression of myogenin (Myog) and myosin heavy chain 2 (Myh2) up-regulated. However, the cell fusion index of myoblasts was higher in grooves with a width of 100 μm, 200 μm and 300 μm. Furthermore, Cytochalasin D disrupted intracellular stress fibers and myotube formation. However, there were still many myotubes formed in the 200 μm groove with the addition of Cytochalasin D. The expression of fusion related genes was higher than the flat group. Moreover, incorporating groove structure into scaffolds of soybean protein isolate and pea protein isolate could enhance cell adhesion, activity and myogenic differentiation of myoblasts. This could provide ideas for scaffolds design of cultured meat products using groove structure.