Design of edible whey protein isolate hydrogels with cell adhesion via a two-step crosslinking method for cultured meat scaffolds

Abstract

Cultured meat is a promising solution to address the global food crisis, with the design of edible scaffolds being a key challenge in its production. Traditionally, materials containing RGD cell adhesion sites, such as gelatin, have been considered necessary to satisfy the need for cultured meat scaffolds with high cell adhesion. However, in this study, we propose a different view in which the biopolymer without inherent cell adhesion sites can also be used as the raw material to prepare edible hydrogel scaffolds for the production of cultured meat, eliminating the need for gelatin or unsafe chemical modifications. We developed edible hydrogel scaffolds from whey protein isolate (WPI) using a two-step crosslinking method to adjust their physical and structural properties, thereby imparting cell adhesive properties. Compared with the single-crosslinked WPI hydrogel, the obtained double-crosslinked WPI hydrogels exhibited higher Young's modulus (∼12.44–25.87 kPa), higher wettability (contact angle <40°), and positively shifted zeta potential, as well as denser gel networks and better hydration properties. These optimized hydrogels supported the adhesion, rapid proliferation, and successful differentiation of mouse skeletal C2C12 myoblasts and porcine muscle stem cells (PMuSCs), with the MTLC hydrogel (crosslinked using 80 U/g TGase and 2 % CaCl₂) showing the best performance. Furthermore, textural analysis revealed that the products we produced were similar to fresh pork in texture. These findings highlight the potential of biopolymers without inherent adhesion sites as viable materials for the development of edible and functional scaffolds in cultured meat production.