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Induced pluripotent stem cells (iPSCs) have become a useful in vitro disease model and a potential therapeutic cell source for ex vivo gene therapy due to their unlimited proliferation capacity and multidirectional potential. To avoid invasive sampling, several methods have been developed to generate human iPSCs from urine-derived cells. Classical reprogramming methods require transfection of c-Myc; however, c-Myc increases the risk of tumor formation, and, therefore, iPSCs that are free of the tumorigenic risk factor c-Myc would be more suitable and safer for basic research and future clinical applications. This chapter details a method for generating c-Myc-free and non-integrating iPSCs from urine cells via an episomal iPSC reprogramming vector, which replaces c-Myc with hsa-miR302–367cluster