Effects of flaxseed protein isolate on myoblast differentiation under low-serum conditions: implications for cellular agriculture.

Background: Oilseed cakes are by-products of oil extraction and they have considerable potential in novel food technologies due to their low cost and high nutrient content. In vitro production of skeletal muscle cell biomass is an emerging approach in cultured meat research and offers a promising alternative protein source. One of the major barriers to large-scale production is the reliance on animal serum, an expensive, animal-derived component. To address this limitation, this study explored the potential of flaxseed protein isolate (FPI), extracted from flaxseed oil cake, as a supplement in cell culture media with a fourfold reduction in serum for the cultivation of C2C12 myoblasts. Results: Flaxseed protein isolate was evaluated for its impact on cell proliferation and differentiation in low-serum media. As the media component, FPI did not enhance C2C12 cell proliferation, but it facilitated myogenic differentiation significantly. Specifically, supplementing low-serum media with 0.25 mg mL^-1 FPI supported cell survival throughout the 10 day differentiation period, resulting in typical myogenic marker presence. It also promoted the formation of myotubes with diameters and fusion index comparable with those observed in control cell cultures with standard serum level. Flaxseed protein isolate also considerably increased the production of sulfated glycosaminoglycans (sGAGs), an extracellular matrix component, exceeding even the levels observed in the control group. Conclusion: The results of this study suggest that flaxseed protein may serve as a valuable supplement in low-serum media to support myoblast differentiation. This could help reduce the cost of cultured meat production while supporting circular economy principles and the reuse of agricultural byproducts.