Harvest of quality-controlled bovine myogenic cells and biomimetic bovine muscle tissue engineering for sustainable meat production.

Alternative technology for meat production holds the potential to alleviate ethical, environmental, and public health concerns associated with conventional meat production. Cultured meat produced using cell culture technology promises to become a viable alternative to animal-raised meat for the future of the food industry. In this study, biomimetic bovine muscle tissue was artificially fabricated from myogenic cells extracted from bovine meat. Our primary culture method relies on three key factors; a sequential digesting process, enzymatic treatment with pronase, and coating with laminin fragment on culture dishes. This method allows the efficient collection of large numbers of primary cells from bovine cheek meat, purifies the myogenic cells from the cell mixture, and then continuously grows the myogenic cells in vitro. In addition, using our "quality control" methods, we were able to determine the "cell quality", including the proliferative and differentiation capability in each step of the primary culture. Furthermore, to mimic native bovine meat, the quality-controlled bovine myogenic cells were cultured on a micropatterned thermoresponsive substrate stimulating a native-like aligned structure of cells, which were then transferred onto a fibrin-based gel. This gel-based culture environment promoted structural and functional maturation of the myogenic cells, resulting in the production of bovine muscle tissues with sarcomere structures, native-like membrane structures, and contractile ability. We believe that these biomimetic features of "tissue-engineered meat" are important for the production of future cultured meat, which will need native-like nutrients, texture and taste. Therefore, our meat production approach will provide a new platform to produce more native biomimetic tissue-engineered meat in the near future.