Human neural stem cells (NSC) are multipotent progenitor cells with the ability to differentiate into several cell types, such as neurons, oligodendocytes, and astrocytes. There is great interest in NSCs as a source for specialized cells for basic neuroscience, drug discovery applications, and cell based therapies for a variety of neurodegenerative diseases including Alzheimer’s, Huntington’s, and Parkinson’s. Neural stem cells can be expanded in vitro either as adherent monolayers or neurospheres. Adherent NSC cultures require pre-coating of culture surfaces with biological materials, such as laminin or CELLstart™ to enable cell adhesion in serum-free conditions. These biological materials have limited shelf life, lot-to-lot variability and the potential for contamination with adventitious agents. To address the limitations of biological growth substrates, Corning Life Sciences, in collaboration with Geron Corporation, developed a novel, xeno-free, synthetic surface, the Corning® Synthemax™ Surface, for stem cell culture applications. This surface is comprised of an acrylate polymer, functionalized with a short peptide sequence derived from the vitronectin protein, to mimic biological ligands for cell adhesion. Our earlier data demonstrated successful long-term self-renewal of multiple human embryonic stem cell lines on Synthemax Surface in xeno-free media. In this study, we describe the application of the Synthemax Surface for the long-term culture and differentiation of ReNcell VN neural progenitor stem cells in serum-free medium. Cell performance on Synthemax Surface was compared to cells on freshly coated laminin surface. Our results demonstrate efficient expansion of NSC on Synthemax Surface over the course of nine serial passages (40 days), with stable doubling time of 32-47 hrs. Cells cultured on Synthemax Surface yielded similar cell number, viability, and morphology to cells cultured on the freshly coated laminin control. Importantly, cells retained normal karyotype, high levels of phenotypic markers (nestin and musashi) and ability to differentiate into neurons and astrocytes after long-term multi-passage culture on Synthemax Surface. Interestingly, cells grown on laminin, but not on Synthemax Surface, began to show a steady decline in nestin/musashi positive cell population around passage 7, indicating more permissive conditions for long-term expansion of multipotent NSC population on Synthemax Surface. Our results suggest that Synthemax Surface, in combination with serum-free medium, provides a complete solution for expansion and differentiation of NSC under xeno-free, defined culture conditions. We believe that Synthemax Surface will be useful for both research purposes and therapeutic applications of NSC.