Abstract
This study explores the feasibility of microcarrier-seeded muscle cell expansion and differentiation in simulated microgravity (µG) conditions, aiming to develop a proof-of-concept for producing cultivated meat in space. Gelatin microcarriers supported C2C12 expansion and myogenesis in static culture. The microcarriers were cultured in 1G (stirred bioreactor) and simulated µG bioreactors. µG slowed down the cell expansion, while the 1G condition showed a significant increase in cell expansion. Cells exhibited nuclear elongation and extended cell bodies over 7 days in both 1G and µG conditions. Flow cytometry and real-time polymerase chain reaction (RT-PCR) revealed enhanced myogenesis in both 1G and µG, though differentiation was delayed and gene expression significantly lower under µG. These results suggest that while µG initiates differentiation, the process is primarily limited to early stages. Despite the slower myogenesis, it remains feasible, and future research should focus on culture conditions to enhance muscle cell functionality in µG.