Desenvolvimento de tecidos musculares espessos para carne cultivada utilizando nanofibras de acetato de celulose randômica

Abstract

Cultured meat is a promising and emerging technique that aims to produce meat for human consumption, using tissue engineering techniques. Cultured meat can be generated by the growth of animal cells in scaffolds, which are biomaterials capable of mimicking the extracellular matrix (ECM). Currently, meat produced using tissue engineering techniques does not have the morphology, ultrastructure and level of differentiation found in native muscle. In this work, we investigated the use of cellulose acetate nanofibers (NAC), in aligned (NACa) and random (NACr) configurations, for the growth and differentiation of muscle lineages. We demonstrate that a scaffold formed by a mesh of randomly distributed fibers (NACr) is an excellent substrate for muscle tissue engineering and, therefore, to produce cultured meat. Our results showed that these matrices not only allow the adhesion and colonization of C2C12 and H9c2 myoblasts, but also trigger their spontaneous myogenesis, without the need for differentiation medium. We demonstrated that the induction of differentiation in these substrates was related to mechanotransduction events, involving signaling from the Hippo-YAP/TAZ pathway. Furthermore, we have successfully developed a new process to stack layers of NACr with cells, enabling the production of a thick meat product. NACr sheets loaded with C2C12 and H9c2 were stacked in up to four layers to form a ~300–400 μm thick and 2 cm long tissue, without compromising the viability of cells within the construct. These results highlight the potential of NACr as an effective substrate in tissue production for cultured meat, enabling future applications with diverse species.