Abstract: Chicken skeletal muscle satellite cells were obtained by a two–step method of collagenase-I and trypsin digestion, followed by in vitro culture. They were further identified by specific gene expression assays. In addition, cell biological characteristics and myogenic genes expression were analyzed. Results showed that satellite cells from chicken skeletal muscle tissue expressed pax7, desmin, and possessed ability to proliferate and differentiate. The technical platform was established to study the mechanism of myogenic proliferation, differentiation and muscle regeneration.