Pleurotus ostreatus and Mucor sp. as alternative cell culture medium ingredients for cellular food production

Fungal extracts in cell culture media offer a sustainable solution to protein limitations in cellular agriculture. With their rich nutritional composition, they provide bioactive compounds supporting cell adhesion, differentiation and matrix formation. In this study, the protein content of Pleurotus ostreatus (PL) and Mucor sp. (M) mycelia was quantified, and their antioxidant (ABTS, FRAP) and ACE inhibitory capacities were initially assessed. Subsequently, the effect of the aqueous extracts from mycelia was evaluated on proliferation and differentiation of C2C12 muscle cells. C2C12 cells were cultured for 24 and 48 hours, during which fungi extracts were administered at different concentrations (from 12.50 to 0.01 mg/ml). Subsequently, cell differentiation was evaluated using the two most effective PL mycelium aqueous extract concentrations (PL1, PL2). After 48 hours of proliferation, different treatments were administered for six days as follows: fetal bovine serum (FBS) + horse serum (HS); PL1 + HS; PL2 + HS. Cell proliferation was assessed using the Alamar Blue assay. PCR was performed to analyze genes involved in proliferation (Cyclin B1 and D1, CDK4) and differentiation (Myh2, MyoG, MRF4, Desmin). Protein content of the two mycelia was 21.50% for PL and 19.80% for M. PL mycelium exhibited antioxidant capacity (243.97±18.03mg Trolox Equivalent/100g and 61.94±1.96mg Acid Ascorbic Equivalent/100g for ABTS and FRAP respectively). PL ACE inhibitory capacity was 38.90±4.57%. PL extract enhanced C2C12 proliferation at concentrations from 0.8 to 0.1mg/ml. These findings suggest that fungi extracts, rich in bioactive compounds, could enhance C2C12 growth.