Neuronal cell cultures offer a crucial tool to mechanistically analyse regeneration in the nervous system. Despite the increasing importance of zebrafish (Danio rerio) as an in vivo model in neurobiological and biomedical research, in vitro approaches to the nervous system are lagging far behind and no method is currently available for establishing enriched neuronal cell cultures. Here we show that magnetic-activated cell sorting (MACS) can be used for the large-scale generation of neuronal-restricted progenitor (NRP) cultures from embryonic zebrafish. Our findings provide a simple and semi-automated method that is likely to boost the use of neuronal cell cultures as a tool for the mechanistic dissection of key processes in neuronal regeneration and development.