SustainableAlternatives to Fetal Bovine Serum: Evaluatingthe Role of Plant and Insect Protein Isolates in Serum-Free Mediafor Bovine Satellite Cell Proliferation in Cultivated Meat Production

This study explores the development and efficacy of serum-free media supplemented with various plant and insect protein isolates as a sustainable alternative to fetal bovine serum (FBS) for cultivated meat production. Focusing on the proliferation and differentiation of bovine satellite cells (BSCs), we tested proteins derived from various plants, insects, and legume byproducts. Our results demonstrated that alfalfa, chickpea cooking liquid, tofu whey, and mung bean proteins at lower concentrations (0.05–0.25 mg/mL) significantly enhanced cell proliferation, with mung bean media outperforming the others. Differentiation assays showed that cells cultured in serum-free media supplemented with mung bean protein maintained their myogenic potential, forming well-differentiated myotubes similar to those observed in FBS-containing media. Live/dead imaging and metabolic profiling further supported these findings, showing healthy cell morphology and active metabolism in the optimized protein media. However, higher protein concentrations resulted in cytotoxic effects. This research highlights the potential of plant protein isolates as key components in serum-free media, providing an ethical, cost-effective, and sustainable media for cultivated meat production. While insect-derived proteins were not effective in supporting cell growth, optimizing their extraction and concentration may enhance their future applicability. Further refinement and long-term studies are needed to improve media composition and scalability for commercial use.