Dissociated cell cultures from fetal mouse hypothalamus patterns of organization and ultrastructural features

Abstract

Dissociated fetal hypothalamic cells mainly taken from 14 day-old mouse fetuses were grown in vitro for increasing time (9 to 60 days). Soon after inoculation the cells partly reaggregated and attached. The small reaggregates were then interconnected by fibers bundles. After the first week the cultures were composed of a continuous basal monolayer of flat and transparent cells, over which various types of refractile cells were lying in discontinuous areas. The ultra-structural study enabled us to identify these cell types, to describe their spatial relationships, and to follow their evolution with time in culture. The basal cell formed several superimposed layers. With increasing age, they displayed typical features of astrocytes and of ependymal cells. The latter exhibited rhythmic ciliary movements in culture. The overlying cells corresponded to three types which were associated in small clumps: primitive neuro-epithelial cells, maturing as well as mature neurons and typical neurosecretory cells. The latter cells were found as early as 9 days of culture of 14 day-old fetal hypothalamic cells and retained their typical features up to two months. Neuronal processes formed very dense networks at the surface of the cultures and terminated within the basal layers. Axon and dendrites were precociously found and were still present after two months. Within axon terminals dense-core vesicles appeared at the same time as neurosecretory cells. Synaptic vesicles and synaptic junctions were found later on.