Specialized cell culture media is a foundational tool for researchers working in diverse areas, from basic and applied research to biopharmaceutical applications. Thermo Fisher Scientific offers media systems for the culture of human and rodent (primary) neural cell types and more recently has focused on identifying conditions that drive stem cell differentiation toward specific neural lineages. AIM: To develop new cell culture systems that enable robust differentiation of human pluripotent stem cells (PSCs) to distinct neuronal subtypes. METHODS: We have adopted a multifaceted approach for driving PSC to neuronal differentiation- 1.Disconnecting specification/regionalization studies from maturation, allowing experimental time line compression and enabling parallel development activities. 2. Utilizing complex Design of Experiment (DOE) approaches and mathematical modeling paired with validated endpoint assays; 3. Incorporating small molecule chemical library screening to identify compounds with desired properties. RESULTS: We demonstrate the feasibility of distinguishing PSC specification from neuronal maturation by utilizing banks of neural stem cells (NSCs), produced in 7 days using GIBCO® Neural Induction Medium. The NSCs provide a good model to screen and optimize conditions driving neural differentiation and maturation. Additional results from definitive and screening DOEs as well as modeling predictions are described. CONCLUSIONS: In the last several years significant advances in stem cell biology have enabled broader adoption of these cells and provided deeper insight into the mechanisms which regulate their growth and specific cell fate determination. In this work we present our approach to harness this insight to develop next generation culture systems to create useful neuronal cell models from PSCs.