3D‐Printed Prolamin Scaffolds for Cell‐Based Meat Culture

Abstract

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.

Review in edible materials for sustainable cultured meat: scaffolds and microcarriers production

“…Also, Koranne et al (2022) used soy lecithin for the 3D printing of hydrolysed collagen in order to prepare a printable edible ink. Cereal prolamins have demonstrated compatibility with muscle cells and have been used to create edible scaffolds (Su et al, 2023), and bread-derived scaffolds with controllable porosity and mechanical properties have also been developed (Holmes et al, 2022). In addition, DaNAgreen Protinet-P is an edible scaffolding product made from isolated plant proteins that did not provide more information (DaNAgreen, 2023).…”

Sculpting the future of meat: Biomaterial approaches and structural engineering for large‐scale cell‐based production

“…Zein protein from barley and rye have been used to develop a cereal prolamin ink for 3D printing and produce a fibrous scaffold which allows for the attachment and proliferation of C2C12 and porcine skeletal muscle satellite cells (Su et al, 2023). This is the first cell-based meat concept that uses a plant-based source for scaffold bioprinting and will motivate more studies investigating sustainable biomaterials for bioprinting.…”

3D Porous Edible Scaffolds from Rye Secalin for Cell-Based Pork Fat Tissue Culturing

“…To address the hurdle, secalin, from rye, was usually incorporated evidenced by the improved flexibility and stretchability, which were attributed to higher molecular weight and longer peptide chains compared to zein. 6 The superior self-assembly behaviors also render secalin a promising scaffolding material. Moreover, the diverse types of prolamins enable the fabrication of scaffolds with appropriate stiffness for soft tissue because the molecular weight of proteins affects the mechanical properties of scaffolds.…”