Scalable production of muscle and adipose cell-laden microtissues using edible macroporous microcarriers for 3D printing of cultured fish fillets
Xuan Zhou, Hongwei Zheng, Yanchi Wu, Haowen Yin, Xiangzhao Mao, Ningyang Li, Huarong Guo, Yaoguang Chang, Xiaoming Jiang, Qinghui Ai, Changhu Xue
Abstract
Cellular agriculture is a novel platform for addressing the issues of protein scarcity, environmental pressures, and food safety. However, expanding seed cells at a large scale remains a prerequisite for achieving industrial cultured meat production. We here propose the production of large-pore-sized edible porous microcarriers (EPMs) by using NaCl to precisely control ice crystal growth during cryogenic crosslinking. Muscle satellite cells (SCs) and adipose-derived stem cells (ASCs) from large yellow croakers exhibit remarkable adhesion, proliferation, and differentiation on gelatin-based EPMs. Following consecutive expansion, SCs and ASCs densities reach 6.25 × 105 and 5.77 × 105 cells/mL, leading to a 499-fold and 461-fold increase in cell numbers, respectively. Subsequently, the mature microtissues are incorporated into a bioink, thereby enabling successful three-dimensional printing of cultured fish fillets with sensory properties similar to native fish fillets. EPM-based cell expansion and food structuring techniques are promising in facilitating large-scale cultured fish meat production. Cell expansion at large-scale remains a big challenge in the growing field of cellular agriculture. Here, the authors present an approach for the production of muscle and adipose cell-laden microtissues using edible porous microcarriers for 3D printing of cultured fish fillets.