Department of Biomedical Engineering, Technion - Israel Institute of Technology, Haifa, 3200003, Israel.
Aleph-Farms Ltd., Rehovot, 7670609, Israel.
Biomaterials. 2022 May;284:121487. doi: 10.1016/j.biomaterials.2022.121487. Epub 2022 Mar 24.
Cultivated meat harnesses tissue engineering (TE) 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) maturation. Mixtures of pea protein isolate (PPI) and soy protein isolate (SPI) with RGD-modified alginate (Alginate(RGD)) were evaluated as prefabricated mold-based and 3D-printed scaffolds for BSC 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) alone, while allowing unhindered BSC 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 attachment and differentiation. Finally, cellular bioprinting was demonstrated with PPI-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.
培养肉利用组织工程（TE）的概念来创造可持续的、可食用的肌肉组织，以满足不断增长的肉类产品需求及其带来的全球影响。由于 3D 打印是一种制造厚而复杂结构的有前途的方法，因此我们的工作主要评估了两种富含植物蛋白的支架成分，作为牛卫星细胞（BSC）成熟的 3D 可打印平台。豌豆蛋白分离物（PPI）和大豆蛋白分离物（SPI）与 RGD 修饰的藻酸盐（Alginate（RGD））的混合物被评估为用于 BSC 培养的预制模具基和 3D 打印支架，最终也被评估为潜在的细胞打印生物墨水。与单独使用 Alginate（RGD）相比，基于模具的富含蛋白质的支架表现出更高的稳定性和刚度，同时允许 BSC 不受阻碍地扩散和成熟。然后使用可食用的、可移动的琼脂支撑浴开发了这两种成分的基于挤出的 3D 打印技术。成功制造的具有明确定义几何形状的构建体支持 BSC 的附着和分化。最后，使用富含 PPI 的生物墨水进行了细胞生物打印。在两种培养配置中观察到打印后的细胞回收，随着时间的推移，细胞活力恢复到约 80-90%。此外，细胞可以在 3D 打印的细胞结构内成熟。由于在我们的支架制造过程中避免了动物衍生材料，并且豌豆蛋白因其低过敏风险而闻名，因此这些发现对进一步的培养肉研究具有很大的前景。
