In vitro neurogenesis, defined as the differentiation of stem cells into functional neurons under controlled laboratory conditions, represents a pivotal tool in contemporary neuroscience research and regenerative medicine. This approach enables precise modeling of neurodevelopmental processes and neurodegenerative disorders, while also supporting the exploration of novel therapeutic strategies. Stem cell–based platforms, including embryonic stem cells (ESCs), induced pluripotent stem cells (iPSCs), and neural stem cells (NSCs), provide versatile and well-characterized sources for generating diverse neuronal lineages. Experimental in vitro models such as neuronal rosettes and neurospheres offer valuable insights into early neural commitment, spatial organization, and lineage specification during neural differentiation. Furthermore, recent advances have highlighted stem cell–derived exosome therapy as a promising cell-free approach for central nervous system (CNS) regeneration. Collectively, this review summarizes major cellular sources, key molecular markers, differentiation mechanisms, and translational applications of in vitro neurogenesis, while also addressing current limitations and outlining future directions in this rapidly evolving field.