Human urine-derived stem cells (hUSCs) are somatic stem cells isolated from fresh urine by the way of centrifugation combined with adherence screening, which are with strong proliferation and multi-differentiation capacity, and have all kinds of similar biological characters as mesenchymal stem cells. HUSCs have the potential of playing an important role in reconstitution of damaged organs and tissues, treatment of the diseases and drug activity and toxicity screening, and urine-induced pluripotent stem cells (u-iPSCs) have been induced by many ways from hUSCs. However, several questions need to be answered prior to developing a technology for drug screen with hUSCs and u-iPSCs, such as the cell source of hUSCs, directional differentiation and efficiency of iPSCs reprogramming. In this review, recent progress in cell source, cell isolation and cultivation, cell biological characteristics, and the application of hUSCs were studied, especially various ways of inducing hUSCs to u-iPSCs and the possible application prospect were summarize, providing a reference for further study and application of hUSCs in drug development and toxicity testing.