Chick embryo extract-driven culture system enhances chicken muscle satellite cell differentiation and 3D myotube formation

The rapid increase in global meat demand has placed significant pressure on traditional meat production. This has led to the emergence of the cultured meat industry as a promising alternative offering sustainable and ethical solutions. This study explored the efficacy of chick embryo extract (CEE) as a potential economic supplement. Specifically, we aimed to identify the optimal culture conditions for chicken muscle satellite cells (CMSCs) using CEE and evaluate their effects on cell proliferation and differentiation. The CEE+horse serum (HS) condition was found to be the most effective for CMSC proliferation, with a significant increase in the expression of myogenic transcription factors. CMSCs cultured in 30% CEE + 3% HS-containing differentiation medium (CEE + 3%HS-DM) showed higher expression of myogenic differentiation markers and formed thicker myotubes compared to the CMSCs cultured in the 5% fetal bovine serum (FBS)-containing differentiation medium (5% FBS-DM). Notably, CMSCs differentiated in CEE + 3%HS-DM displayed consistently aligned myotubes in the predominant direction. In contrast, myotubes in the 5% FBS-DM showed less directional preference. To investigate 3D myofiber formation on plant-based scaffolds, CMSCs were cultured on a decellularized green onion scaffold. CMSCs successfully attached and differentiated into myofibers, with significantly enhanced attachment and differentiation observed in CMSCs cultured in CEE + 3%HS-DM compared to those in 5% FBS-DM. These results demonstrate that CEE + 3%HS-DM is suitable for CMSC differentiation and 3D myotube formation, providing a promising culture system for sustainable cultured meat production.