Cells dissociated from cerebral hemispheres of 6-day-old chick embryos were cultured either in standard nutrient medium or in the presence of a brain extract from 8-day-old chick embryo. Morphological observations showed the development of bipolar and multipolar neurons in both culture conditions and acetylcholinesterase activity was found in all neuronal cells. Brain extract stimulated the morphological maturation of neurons, expressed by the formation of fiber bundles, fine structural maturation and development of synapses rich in clear vesicles. Furthermore, acetylcholinesterase and choline acetyltransferase activities were higher in the cultures treated with brain extract. In these cultures, the values of choline acetyltransferase activity reached a peak at 10 days and then decreased. These observations are discussed with particular reference to proliferation, maturation and degeneration of cholinergic neurons.