Abstract
Owing to the increase in the global demand of meat, cultured meat technology is being developed to circumvent a shortage of meat in the future. However, methods for construction of millimetre-thick bovine muscle tissues with highly aligned myotubes have not yet been established. Here, we propose a culture method for constructing 3D-cultured bovine muscle tissue containing myotubes aligned along its long-axial direction, which contracted in response to electrical stimulation. First, we optimised the composition of biomaterials used in the construction and the electrical stimulation applied to the tissue during culture. Subsequently, we fabricated millimetre-thick bovine muscle tissues containing highly aligned myotubes by accumulating bovine myoblast-laden hydrogel modules. The microbial content of the bovine muscle tissue cultured for 14 days was below the detection limit, indicating that the muscle tissues were sterile, unlike commercial meat. Therefore, the proposed construction method for bovine muscle tissues will be useful for the production of clean cultured steak meat simulating real meat.
Highlights
To construct contractile bovine muscle tissue, we cultured bovine myocyte-laden hydrogel on a culture device consisting of anchors with pillars fabricated using stereolithography and a substrate consisting of polydimethylsiloxane (PDMS) (Supplementary Fig. 1)
Optimisation of appropriate culture condition has been proposed for conventional formation of cultured muscle tissues, the cell types used were limited to mouse, rat, or human[13,17,18,19]; the culture conditions for bovine muscle tissue were not clear
We found that application of electrical pulses combined with the use of fibrin-matrigel enabled contraction of all bovine muscle tissue
Summary
The global consumption of meat is increasing with population growth, leading to concerns of a future protein crisis.[1,2] As the conventional livestock industries have been problematic for sustainability due to the ethical problems and its adverse effects on the environment,[3] more sustainable technologies for meat production are required to bridge the demand-supply gap and thwart the protein crisis.[4,5] Cultured meat constructed via tissue culture of animal cells is one of the promising candidates as sustainable next-generation meat, as it can be generated using small amounts of cells obtained without killing livestock, as well as lower land use and water footprint.[6]Cultured meat has been constructed using 3D culture methods developed in the fields of regenerative medicine and drug discovery. We develop a method for fabricating millimetre-thick bovine muscle tissues containing highly aligned myotubes using bovine myoblast-laden hydrogel modules with striped structures (Fig. 1).
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