Development of a serum‐free medium for myoblasts long‐term expansion and 3D culture for cell‐based meat

Abstract

Cell‐based meat technology provides an effective method to meet the demand for meat, while also posing a huge challenge to the expansion of myoblasts. It is difficult to develop serum‐free medium suitable for long‐term culture and large‐scale expansion of myoblasts, which causes limited understanding of myoblasts expansion. Therefore, this study used C2C12 myoblasts as model cells and developed a serum‐free medium for large‐scale expansion of myoblasts in vitro using the Plackett–Burman design. The serum‐free medium can support short‐term proliferation and long‐term passage of C2C12 myoblasts, while maintaining myogenic differentiation potential well, which is comparable to those of growth medium containing 10% fetal bovine serum. Based on the C2C12 myoblasts microcarriers serum‐free culture system established in this study, the actual expansion folds of myoblasts can reach 43.55 folds after 7 days. Moreover, cell‐based meat chunks were preliminarily prepared using glutamine transaminase and edible pigments. The research results provide reference for serum‐free culture and large‐scale expansion of myoblasts in vitro, laying the foundation for cell‐based meat production.Practical ApplicationThis study developed a serum‐free medium suitable for long‐term passage of myoblasts and established a microcarrier serum‐free culture system for myoblasts, which is expected to solve the problem of serum‐free culture and large‐scale expansion of myoblasts in cell culture meat production.