<p>This study aimed to compare the proliferation, differentiation rates, and related gene expression levels of skeletal muscle satellite cells (SMSCs) isolated from Hanwoo steers and cows to identify a suitable source for cell-based meat production. SMSCs were extracted from three steers and three cows, and their cell growth rates, doubling times, differentiation rates, and genetic differences were determined. Comparative analysis revealed that SMSCs from steers exhibited a higher growth rate than cows. Doubling time was shorter in steers than in cows (p&lt;0.0001). During differentiation, the cell fusion index showed significant differences between steers and cows (p&lt;0.0001). Steers showed differentiation after 24 h, while cows showed differentiation after 72 h. Genetic analysis showed that the expression level of the Pax7 gene was significantly higher in steers than in cows (p&lt;0.001). The expression levels of differentiation-related genes, including MyoG, MRF4, and MHC1 were higher in steers than in cows. Thus, SMSCs isolated from steer muscle exhibit a faster growth rate, and these findings are expected to provide valuable information for determining the optimal cells for cell-based food.</p>