Evaluating the Efficacy of Serum-Free Media Supplemented with Protein Isolates for Bovine Satellite Cell Proliferation: A Sustainable Approach for Cultivated Meat Production

Abstract

This study investigates the short- and long-term efficacy of serum-free media supplemented with protein isolates from algae, alfalfa, silkworm pupae, and grasshoppers for the proliferation of bovine satellite cells (BSCs). Fresh and spent media were analyzed to monitor metabolites, while cell proliferation was assessed using the CyQUANT assay. The results of the short-term growth study indicated that a lower concentration of alfalfa protein isolate (0.05 mg/mL) significantly enhanced cell proliferation, achieving a 1.47-fold increase compared to basal media, thereby demonstrating its potential as a viable alternative to fetal bovine serum (FBS). The capability of the developed serum-free media for cell passaging was examined using different coating strategies, with vitronectin outperforming others; however, it was unable to support the long-term growth of cells compared to FBS-containing media. Given the critical role of glutamine, this study also evaluates the impact of L-glutamine supplementation at different concentrations on cell growth and metabolism in alfalfa-based media. However, while glutamine supplementation showed trends toward increased cell growth, the enhancements were not statistically significant. Based on the results, this study highlights the potential of alfalfa protein isolate as a promising component of serum-free media for BSC proliferation and underscores the need for continued research into alternative protein sources and media formulations to support the sustainable and ethical production of cultivated meat.