Generation of in vitro hiPSCs-derived skeletal muscle progenitor cells (SMPC) holds a great promise for regenerative medicine as it might provide a therapeutic solution to re-build lost myofibers in neuromuscular diseases, including example Duchenne Muscular Dystrophy (DMD). However, hiPSCs-derived SMPC are usually obtained in a low amount and resemble a more embryonal/fetal stage of differentiation. Building on previous discoveries on the key role of JNK/STAT3 pathway in controlling cell-fate decision of SMPC, we hypothesized that pharmacological inhibition of the pathway could increase the population of PAX7+ muscle progenitors. Transient treatment of myogenic cultures with AG490, a JNK/STAT3 pathway inhibitor, could double the number of hiPSCs-derived PAX7+ SMPC and increase the number of muscle stem cell with higher levels of PAX7 expression. Importantly, these SMPCs can maintain the ability to differentiate in vitro upon removal of AG490 from the media. We illustrate an effective procedure for SMPC isolation by CD54 sorting, as a way to obtain a pure population of SMPCs suitable for in vivo transplantation. This procedure was effective using either hiPSC derived from healthy donor, as well as hIPSCs derived from DMD patients with distinct mutations (DMD∆52 and DMD∆8/9), suggesting that JNK/STAT3 inhibition could be used as a universal strategy to generate hiPSCs-derived SMPC for therapeutic purposes.