Apigenin as a media supplement to promote muscle satellite cell differentiation for cultured meat production: an in silico and in vitro study

Muscle satellite cells (MSCs) are essential for cultured meat (CM) production, and natural compound supplementation can enhance their proliferation and differentiation. Thus, this study used in silico analysis to identify natural compounds inhibiting myostatin (MSTN; a negative regulator of muscle development) and promoting MSC differentiation. Apigenin showed stable binding to MSTN (−6.5 kcal/mol) and was confirmed to directly interact with MSTN by cellular thermal shift assay, indicating its potential as a promising compound. Next, myoblasts (C2C12) and MSCs derived from bovine, chicken, and porcine species were cultured in media supplemented with apigenin. This led to increased expression of myosin heavy chain and downregulation of MSTN. Furthermore, the expression levels of SMAD2 and SMAD3, as well as their phosphorylated forms, which are downstream components of MSTN signaling, were also reduced. Notably, chicken MSCs exhibited the most pronounced response to apigenin supplementation, with differentiation reaching 235% and 135% of the control (control set as 100%) at 0.1 nM and 1 nM, respectively. Similar trends of enhanced MSC differentiation were observed in other species at 0.1 nM, with C2C12 cells increasing by 118% and bovine MSCs by 174%. Additionally, apigenin supplementation inhibited the production of intracellular reactive oxygen species (ROS). Therefore, apigenin shows potential as a natural additive or media supplement for CM production by promoting MSC differentiation, inhibiting MSTN, and suppressing ROS activity. Thus, this study suggests a potential strategy for improving the efficiency and quality of CM production through the supplementation of natural compounds. • Apigenin binds and inhibits myostatin (MSTN), promoting muscle cell differentiation. • Supplementation enhanced differentiation of MSCs from chicken, bovine, porcine, and C2C12 cells. • Apigenin reduced intracellular ROS, supporting its use as a natural additive in cultured meat production.