Development of Polysaccharide Hydrogels Enriched With Protein and Starch for Bovine Myoblast Cell Culture

The expanding worldwide population has increased the meat demand, prompting efforts to find alternatives. A promising approach is the cultivation of animal cells on edible biomaterials for cultured meat production. However, those biomaterials face challenges in their mechanical properties, cytotoxicity, and ability to support optimal cell growth. In this study, we focused on optimizing plant-edible hydrogels as a 3D environment for the growth of bovine myoblast cells. We prepared alginate hydrogel (A) to be enriched with soybean protein (S.A) and tapioca starch (T.A), developing Group 1 hydrogels. Aiming to enhance their elasticity, xanthan gum (XG) was incorporated into Group 1, generating alginate-xanthan gum (Ax), soybean-alginate-xanthan gum (S.Ax), and tapioca-alginate-xanthan gum (T.Ax) Group 2 hydrogels. Both groups were assessed for physical and chemical analyses, rheological testing, cell viability assays, immunofluorescence staining, gene expression, and flavor profiling. Our findings showed that all hydrogels maintained their crosslinking for up to 7 days except Ax and T.Ax, which showed degradation of 57.39% and 36.03%, respectively. Both groups represented swelling characteristics, porosity, protein adsorption, and cooking capabilities. Moreover, A and S.A exhibited viscous properties with slow stress relaxation, whereas T.A displayed rapid relaxation and viscoelastic behavior. Successfully, Group 2 demonstrated faster stress relaxation and sufficient elasticity. Bovine myoblast cells showed no significant toxicity and could proliferate, expressing paired box 7 (PAX-7) marker in both groups. At the differentiation stage, desmin expression indicated the intermediate differentiation of the muscle cells for up to 7 days in both groups. Besides S.A and S.Ax scaffolds exhibit the nearest metabolic similarity to beef among the plant-based scaffolds. These findings suggest that the hydrogels enriched with protein and starch holding the potential for culture meat production.