A chemically defined medium has been formulated which supports the growth (proliferation and differentiation) of rat- and ovine-derived myogenic satellite cells in vitro. Utilization of this medium in a direct comparison study in which satellite cells from both species were exposed to insulin resulted in the following observations: (1) insulin promoted the dose-dependent proliferation of primary cultures of ovine-derived but not rat-derived satellite cells and (2) rat-derived satellite cells fused to form multinucleated myotubes when exposed to increasing levels of insulin. Collectively, these observations suggest that the rat satellite cell culture system may not be an appropriate model system for extrapolating in vitro growth data to variables of ruminant skeletal muscle growth.