Mammalian cell behavior is guided by a three-dimensional (3D) microenvironment that provides chemical, physical, and biological cues through interactions between the extracellular matrix (ECM), other cells, and soluble factors. A major challenge lies in recapitulating the complexity of the microenvironment in a systematic way to understand and direct cell fate. Unlocking the promise of human pluripotent stem cell (hPSC)-based therapy necessitates creating precise, scalable cell culture systems that streamline expansion, differentiation, and isolation procedures. Strategies used to maintain stemness and differentiate human pluripotent stem cells (hPSCs) in vitro are predominantly based on two-dimensional (2D) systems, where small molecules and growth factors are key regulators. Recent advances in 3D systems, mainly established on organoid systems, have provided outstanding approaches to studying various diseases and organ developments. Yet, both systems have their own limitations. Here, we leveraged an engineered 3D scalable culture platform that can considerably advance the outcomes of the current platforms by (1) maintaining the undifferentiated state of hPSCs in a defined medium, and (2) directing neural differentiation in the presence of external growth factors in scaffolds. Our engineered 3D culture platform facilitates a reliable method for maintaining stem cell self-renewal and differentiating cells for large-scale cultivation of hPSCs in regenerative medicine and tissue engineering.