Muscle tissue culture provides a system for studying the growth and differentiation of muscle cells in a controlled environment. In mature muscle tissue, terminally differentiated myocytes form multinucleate syncytia in which structural and regulatory genes are expressed and the contractile apparatus is assembled. Adult muscle fibres are characterized by the presence of satellite cells. These are a quiescent population of myogenic cells that reside between the basal lamina and the plasmalemma of terminally differentiated muscle fibres and are rapidly activated in response to appropriate stimuli. This chapter describes protocols used in our laboratory for isolating and culturing satellite cells isolated from mouse skeletal muscles. In particular we discuss the technical aspect of satellite cell isolation, the methods necessary to enrich the satellite cell fraction, and the culture conditions which optimize proliferation and myotube formation of mouse satellite cells.