Morphological and biochemical maturation of neurones cultured in the absence of glial cells

Abstract

Cultures of isolated neurones offer possibilities for investigating their properties directly in the absence of glial cells. Furthermore, such a culture provides a suitable model for the study of the biochemical and functional maturation of neuronal cells in a well defined molecular environment. Culture techniques of isolated neurones have been developed for cells originating from the peripheral nervous system¹, especially sympathetic ganglion cells^2–5. Neurones from the central nervous system (CNS) have proved more difficult to maintain in culture in the absence of other cell types⁶. It is only recently that hippocampal neurones from rat embryos have been successfully maintained in culture⁷. We now describe a method for obtaining a pure culture of dispersed neurones from the telencephalon of 8-day-old chick embryos. This method is derived from a technique previously developed in our laboratory⁸. In those culture conditions, the neuronal cells were obtained by dissociation of cerebral hemispheres from 5–7-day-old chick embryos. The cells remained isolated for only 48 h after plating. Beginning with the 3rd day in vitro, the cells clumped together and formed large aggregates in which it was quite difficult to identify the cell types. We have improved those culture conditions and have succeeded in cultivating neurones which remained well dispersed for the whole period of culture.