The overall goal of this project is to develop a stem cell-based therapy for the treatment of urinary incontinence in women; therapies are also applicable for men. Urinary incontinence is very common and is linked to aging and to complications of pregnancy due to injury of the urethral sphincter. Our strategy consists of deriving induced pluripotent stem cell (iPSC) lines from patients with a non-integrative technology that would not alter the gene content of cells, differentiating these lines to smooth muscle cells, assessing characteristics of undifferentiated and differentiated cells, testing safety and efficacy in animal models, and proceeding to initial discussions with the FDA. We have derived three iPSC lines from patients, characterized the lines for standard properties of stem cell lines, optimized differentiation further and established initial benchmark assays for analysis of undifferentiated cells, as well as, established methods for animal studies. We have also established protocols that eliminate animal cells and proteins from culture and media in order to provide the safest possible cells for eventual cell replacement therapies. In the current funding period, we have concluded two animal efficacy studies using our stem cell-derived therapy. We demonstrated that the therapy consistently restores urethral function in the animal model. We are on track with our milestones and are confident that the project will yield important results for the women, and men, of the state of California. In 2008, 20,330 women underwent surgery for urinary incontinence in California; this extrapolates to 172,500 UI procedures/year nationally (76,500 outpatient and 96,000 inpatient) and more than 100,000 procedures in California over the last 5 years. This number is forecast to increase by 55% by 2050. Yet, although short-term results of surgery are good, long-term results are not. Alternatives such as minimally invasive methods of injecting bulking agents, including collagen or autologous fat, provide only temporary and partial relief. Thus, our project is important as it aims to improve treatments in order to assist the significant number of patients without options for relief in the state of California.