Cultivating meat from muscle stem cells in vitro requires 3D edible scaffolds as the supporting matrix. Electrohydrodynamic (EHD) printing is an emerging 3D‐printing technology for fabricating ultrafine fibrous scaffolds with high precision microstructures for biomedical applications. However, edible EHD‐printed scaffolds remain scarce in cultured meat (CM) production partly due to special requirements with regard to the printability of ink. Here, hordein or secalin is mixed, which are cereal prolamins extracted from barley or rye, with zein to produce pure prolamin‐based inks, which exhibit favorable printability similar to common polycaprolactone ink. Zein/hordein and zein/secalin scaffolds with highly ordered tessellated structures are successfully fabricated after optimizing printing conditions. The prolamin scaffolds demonstrated good water stability and in vitro degradability due to the porous fiber surface, which is spontaneously generated by culturing muscle cells for 1 week. Moreover, mouse skeletal myoblasts (C2C12) and porcine skeletal muscle satellite cells (PSCs) can adhere and proliferate on the fibrous matrix, and a CM slice is produced by culturing PSCs on prolamin scaffolds with high tissue similarity. The upregulation of myogenic proteins shows that the differentiation process is triggered in the 3D culture, demonstrating the great potential of prolamin scaffolds in CM production.

Hydrogel microbeads are promising matrices for cell-based meat culture because they can be conveniently fabricated from food-grade materials. In this study, food-grade sodium alginate was successfully applied to fabricate edible microbeads with the desired diameters (~200 µm) using electrospray technology. Pure alginate microbeads have poor cytoaffinity. To enhance the cytoaffinity of alginate microbeads for meat culture, we applied different gelatin coatings, namely porcine gelatin, bovine gelatin, and fish gelatin, onto alginate beads. The modification is non-toxic and environmentally friendly with the use of plant-derived crosslinkers and under mild reaction conditions. As all ingredients are food grade, the modified microbeads are edible and biodegradable, with the mean diameters of 142.96 ± 35.49 μm (porcine gelatin-coated beads), 105.12 ± 80.46 μm (bovine gelatin-coated beads), and 94.81 ± 67.38 μm (bovine gelatin-coated beads). These modified microbeads have self-fluorescence and can be easily distinguished under a fluorescent microscope. The hydrogel core enables the microbeads to have a density of 1.04 g/cm3 for suspension culture. Bovine gelatin-coated microbeads have the best primary porcine myoblast and C2C12 cell adhesion, and can achieve between 16-to 18-fold proliferation of different muscle cells. With the successful differentiation of C2C12 and proliferation of 3T3-L1 adipocytes on bovine gelatin-coated microbeads, cells and microbeads can form clusters and become spheroids, illustrating that our edible microbeads are a promising microcarrier for mammalian cell culture for cell-based meat production.

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