Phenotypic diversity among mature skeletal muscle fibers originates from muscle progenitor cells, primary and secondary myoblasts, each of which is intrinsically committed to express a characteristic complement of developmentally regulated myosin heavy chain genes when differentiated. Similarly, postnatal muscle myoblasts, the satellite cells nestling beneath basement membranes of mature skeletal muscle fibers, have been shown to exhibit diversity, related to whether the muscle in which they reside is of a slow, fast or superfast type. Here we analyzed this association in more detail, evaluating the myosin heavy chain gene expression in immature muscle fibers (myotubes) formed in vitro from satellite cells extracted from isolated, living, single muscle-fibers of mature murine muscle. We identified a population of satellite cells that form myotubes expressing type I (slow) myosin heavy chain and found this population to be preferentially associated with individual slow muscle-fibers. These results not only confirm diversity among mammalian satellite cells, but also demonstrate that the phenotype of satellite cells is indicative of the type of fiber from which they derive.