Induced pluripotent stem cells obtained by reprogramming of primary somatic cells have revolutionized the cell biology and disease modeling fields. However, modeling human skeletal muscle and neuromuscular disorders has been hindered by a limited number of protocols for generation of mature muscle fibers with sarcolemmal organization. Through simultaneous co-differentiation of hiPSC into muscle cells and motor neurons, we developed a novel procedure for generating innervated multinucleated mature skeletal muscle fibers. Presence of both cell types greatly enhances myoblast differentiation and yields mature functional millimeter-long multinucleated muscle fibers. Furthermore, this organoid-like culture can be maintained over long periods of time with autonomous cell regeneration thanks to the presence of PAX7-positive cells and extracellular matrix synthesis. This protocol applicable to hiPSCs from healthy individuals was validated in Duchenne Muscular Dystrophy, Myotonic Dystrophy, Facio-Scapulo-Humeral Dystrophy and type 2A Limb-Girdle Muscular Dystrophy opening new paths for exploration of muscle differentiation, disease modeling and drug discovery.