The aim of this study was to develop a technique to culture satellite cells from isolated intact fast or slow human muscle fibers. Previous studies have been carried out on small rodent muscles where the fibers run from tendon to tendon, but this is the first description of the modification of this technique for much larger human muscles. We have demonstrated that the human muscle fibers are in fact segmental, and we have also shown that it is possible to obtain very pure satellite cell cultures. We discuss the importance of this technique as a source of highly purified muscle cell cultures, which can be used for further studies on satellite cell behavior.