Porous edible cryogel inspired by the production of shimitofu (freeze-dried tofu) for high-density cell culture in cultured meat

Abstract

The increasing global demand for alternative proteins has driven research efforts toward the development of innovative and energy-efficient food production methods. This study focuses on developing a manufacturing technology for mature tissues that mimic the physical and biochemical properties of biological tissue using edible porous hydrogel materials derived from soy protein as three-dimensional culture scaffolds. We investigated a combination of soy protein isolate (SPI), known for its excellent cell adhesion properties, with carrageenan (CA) and sodium alginate (SA), both of which are known for their rapid ion-cross-linking capabilities. In the experiments, SPI, CA, and SA were dissolved in water, heated, mixed, and then allowed to gel at room temperature. Inspired by the production process of shimitofu , a traditional Japanese freeze-dried tofu, the hydrogel was subjected to repeated heating and cooling to enhance gelation, after which it was immersed overnight in Ca 2+ and K + solutions to form SPI/CA/SA hydrogels. A cryogel was subsequently produced by freeze-drying. The cryogel when seeded with myoblasts (C2C12) and cultured under agitation exhibits improved mechanical strength, with significant cell adhesion and proliferation observed after 10 days. These findings suggest that the porous structure promotes cell growth, highlighting the biocompatibility of the SPI/CA/SA cryogel. Therefore, it is an effective scaffold for cultured meat production and offers a sustainable solution for protein production. • A porous, anisotropic cryogel scaffold, inspired by the structure of the traditional Japanese food shimitofu , has been developed for cultured meat applications using soy protein isolate (SPI), carrageenan (CA), and sodium alginate (SA), without the need for chemical cross-linking agents. • All materials used in the SPI/CA/SA scaffold are fully edible, an essential requirement for cultured meat products. • The SPI/CA/SA cryogel scaffold features a highly porous structure with interconnected pores, enhancing cellular infiltration and improving nutrient and oxygen diffusion. • The anisotropic structure of the SPI/CA/SA scaffold facilitates muscle fiber alignment and formation, which is critical for replicating the organization of natural tissues.

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