Advances in human pluripotent stem cells (hPSCs) research have provided the potential hope for significant improvements of disease treatment and management. The success of stem cell-based therapy will have major impacts on the quality of life of people with chronic health problems such as cancer, cardiovascular diseases, and neurodegenerative disorders. The realization of the full potential of hPSCs in regenerative medicine requires, among other things, the establishments of well-defined culture conditions for their growth and differentiation and cost-effective protocols for their expansion. In this grant application, we propose a series of experiments to develop a novel technology platform using cost-effective and well-defined synthetic matrices to expand and differentiate hPSCs. During the funding period, we developed a comprehensive approach to elucidate the responses of hPSCs, as well as neuronal progenitor cells (NPCs) to microenvironmental factors in a combinatorial and systematic manner. Application of this novel and powerful technology will lead to the definition of the optimal synehtic matrices for the control of hPSC growth and differentiation and the production of hPSCs and NPCs without contamination by non-human products.