Skeletal muscle contains both differentiated myofibers and stem cells, known as satellite cells. The satellite cells, comprising around 1% of the total muscle nuclei, are present between the plasma membrane of the multinucleated muscle cells and the basal lamina that surrounds each myofiber. In adult muscle, satellite cells are quiescent but proliferate in response to muscle injury, producing myoblasts that can either form new satellite cells or fuse with one another or pre-existing multinucleated muscle cells. This process help to repair the muscle. They are responsible for postnatal muscle growth, hypertrophy and regeneration of skeletal muscle. Upon quiescent satellite cells activation, they co-express the transcription factors Pax7 and myoD. In vitro culture of Human Skeletal Muscle Satellite Cells is a useful model for studying the process of cell differentiation. They are also useful tools to stablish in vitro disease models for High Throughput and High Content Screening.