Functional and nutritional evaluation of Pleurotus ostreatus for food and cellular agriculture applications

The increasing global demand for sustainable protein sources calls for innovative biotechnological strategies to reduce reliance on animal-derived ingredients. In this context, the present study explored Pleurotus ostreatus as a promising fungal matrix for food and cellular agriculture applications. The objective was to assess its nutritional composition, antioxidant profile, and potential use as a functional component in cell culture media. Fungal biomass (FB) obtained through liquid submerged fermentation was characterised for its nutritional profile and functional activity after in vitro digestion (Infogest 2.0) in comparison with whey (WH) and yeast (YT); alternatives already used in food sector. Furthermore, a 100% aqueous extract of P. ostreatus (PL) submerged mycelium was tested on murine skeletal muscle cells (C2C12), as alternative to animal-origin ingredient in cell culture media, to evaluate its effects on viability, morphology, and gene expression. Results showed that fungal mycelium exhibited moderate protein content (21.33 ± 0.21 g/100 g), high fiber levels (19.05 ± 0.42 g/100 g), and notable antioxidant activity, with significantly higher phenolic content than YT. Despite lower digestibility than WH, FB demonstrated strong antioxidant potential after digestion, confirming the presence of bioactive compounds. In cell-based assays, PL enhanced C2C12 viability and proliferation at 48 h, particularly when combined with insulin–transferrin–selenium (ITS) and bovine serum albumin (BSA), resulting in improved morphology and upregulation of genes related to proliferation and differentiation (Cyclin D1, CDK4, MRF4). These findings indicate that FB can act as a sustainable, functional ingredient for both food systems and serum-reduced media in cultivated meat production, supporting a transition toward more ethical and circular protein production models.