Exploring Plant-Based Proteins as an Alternative for Culture Media in Cellular Agriculture

The food sector is currently one of the most debated fields, particularly regarding its sustainabil-ity. While the livestock industry has actively worked to mitigate its environmental impact, scien-tific innovation must contribute further to address this challenge. In this context, cultured meat emerges as a revolutionary option, with the potential to overcome many ethical and environmen-tal issues associated with conventional meat production. However, the development of this inno-vative product faces significant challenges, chief among them being the formulation of culture media that are free of animal-derived components, economically sustainable, and compatible with cellular requirements. For this reason, the aim of this work was to evaluate protein extracts de-rived from hempseeds (HSs) as constituents of the culture medium for murine C2C12 muscle cells. Specifically, following alkaline extraction and isoelectric precipitation, protein isolates from HSs were tested at various concentrations (1:2 dilutions) on C2C12 cells. The results demonstrat-ed that all concentrations, except for the initial stock solution, effectively supported cell growth and proliferation for up to 48 h, as assessed using the Alamar Blue colorimetric assay. Among all the tested concentrations, 0.6 mg/mL was selected for further analysis and incorporated as a component of an edible bioink used for 3D bioprinting. The experiments showed that 0.6 mg/mL of HS protein extract was capable of sustaining cell viability in 3D structures for up to 7 days, as confirmed by the Live/Dead assay and confocal microscopy analysis. These findings indicate that HSs, characterized by high nutritional profile (94.6±0.12% of DM; 23.1±0.57% of crude protein; 27.9±0.75 of lipids), hold great potential as a sustainable, accessible, and alternative protein source for the formulation of culture media in the context of cellular agriculture. Further research is required to explore other plant-based matrices or food industry co-products that could reduce costs and optimize cell growth parameters. The integration of natural-origin ingredients with ad-vanced technologies such as bioprinting could pave the way for novel approaches to cultivated meat production, bringing this solution closer to commercial reality and aligning it with the prin-ciples of a circular economy.