Updated project metadata. An organized network of beating cilia transports cerebrospinal fluid (CSF) through the ventricles of the brain. CSF takes along diverse solutes including extracellular vesicles (EV), which arise from the choroid plexus, a secretory epithelium that reaches into the ventricles. Along their path through the ventricles, EVs have the opportunity to contact neural stem cell (NSC) niches. We purified EVZ310 produced by a choroid plexus-cell line, by differential centrifugation and flotation in iodixanol density gradients. We co-cultured suspensions of EVZ310 with NSC from the lateral and third ventricle. Alternatively, EVZ310 were dried-down on a polymer substrate and a suspension of NSCs was added. In both cases, EV Z310 induced, in a dose dependent manner, the NSC to rapidly form cellular networks accompanied by the expression of genes characteristic for neurons (Tuj1) and astrocytes (Gfap). NSCs from either origin responded to EVZ310 qualitatively and quantitatively in a very similar way. By contrast, EVMEF purified from mouse embryonic fibroblasts had little effect on both types of NSC. Mass spectrometry revealed significant differences in composition between EVMEF and EVZ310 proteomes. Notably, EVZ310 were enriched for the membrane or membrane associated proteins known to be involved in neurogenesis, cell differentiation, membrane trafficking and membrane organization.