A serum-free media formulation for cultured meat production supports bovine satellite cell differentiation in the absence of serum starvation [Timecourse]

Production of cultured meat requires the robust differentiation of satellite cells into mature muscle fibers without the use of animal-derived components. Current protocols induce myogenic differentiation in vitro through serum starvation, an abrupt reduction in serum concentration. Here, we used RNA sequencing to investigate the transcriptomic remodelling of bovine satellite cells during myogenic differentiation induced by serum starvation. We characterized canonical myogenic gene expression, and identified surface receptors upregulated during the early phase of differentiation. Supplementation of ligands to these receptors enabled the formulation of a chemically defined media that induced differentiation in the absence of serum starvation and/or transgene expression. Serum-free myogenic differentiation was of similar extent to that induced by serum starvation, as evaluated by transcriptome analysis, protein expression and the presence of a functional contractile apparatus. Moreover, the serum-free differentiation media supported the fabrication of mature three-dimensional bioartificial muscle constructs, demonstrating its suitability for cultured beef production. Satellite cells were cultured in growth media (GM) containing 20% FBS and abuptly switched to differentiation medium containing 2% FBS, a procedure known as serum-starvation. RNA samples were taken at 0 h, 24 h, 48 h, 72 h and 96 h of serum starvation in biological quadruplicates (A, B, C, D).