3D-printable plant protein-enriched scaffolds for cultivated meat development

Cultivated meat harnesses tissue engineering ([TE]{.caps}) concepts to create sustainable, edible muscle tissues, for addressing the rising meat product demands and their global consequences. As 3D-printing is a promising method for creating thick and complex structures, two plant-protein-enriched scaffolding compositions were primarily assessed in our work as 3D-printable platforms for bovine satellite cells ([BSC]{.caps}) maturation. Mixtures of Pea protein isolate ([PPI]{.caps}) and soy protein isolate ([SPI]{.caps}) with [RGD]{.caps}-modified alginate (Alginate ([RGD]{.caps})) were evaluated as prefabricated mold-based and 3D-printed scaffolds for [BSC]{.caps} cultivation, and ultimately, as potential bioinks for cellular printing. Mold-based protein enriched scaffolds exhibited elevated stability and stiffness compared to ones made of Alginate ([RGD]{.caps}) alone, while allowing unhindered [BSC]{.caps} spreading and maturation. Extrusion based 3D-printing with the two compositions was then developed, while using an edible, removable agar support bath. Successfully fabricated constructs with well-defined geometries supported [BSC]{.caps} attachment and differentiation. Finally, cellular bioprinting was demonstrated with [PPI]{.caps}-enriched bioinks. Cell recovery post-printing was observed in two cultivation configurations, reaching ∼80--90% viability over time. Moreover, cells could mature within 3D-printed cellular constructs. As animal-derived materials were avoided in our scaffold fabrication process, and pea-protein is known for its low allergic risk, these findings have great promise for further cultivated meat research.