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 ultrastructural 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.

Expression of Peptides Derived from the Melanin-Concentrating Hormone Precursor in Serum-Free Culture of Rat Fetal Hypothalamic Neurons: Role of Attachment Factors

“…Our results demonstrate that primary cultures of dis sociated cells from fetal posterior hypothalamus can be maintained for more than 1 month under SFM condi tions, as already shown by other groups in other neuron cultures [18,22,23].…”

“…Matrigel favored the formation of astroglial cell clusters linked with 'cables'. According to the studies made on dissociated fetal cells from different brain areas [20,22], this aspect is similar to that observed when cerebral hemisphere cells were cultured.…”

Immunofluorescent Patterns of Dissociated Rat Embryo Cerebral Cells during Development in Surface Culture: Distinctive Reactions with Neurite and Perikaryon Cell Membranes

“…Aggregation under culture conditions appears to be an essential step for the establishment of inter cellular cooperativity and may be largely re sponsible for the ability of cells to express their functional properties [Seeds, 1971;God frey et al, 1975], In a previous study we have shown that the aggregation process in surface brain cell cultures predates the appearance of axonal and dendritic extensions as well as the establishment of synaptic contacts and forma tion of myelin [Yavin and Yavin, 1977]. While a great deal of experimental evidence has been accumulated on the histotypic organiza tion of cells in suspension cultures [Garber and Moscona, 1972a,b], little information is presently available as to the cell types com prising the cerebral aggregate as it evolves under surface culture conditions [Benda et a!., 1975].…”

Differentiation of Fetal Mouse Hypothalamic Cells in Serum-Free Medium

“…It is shown here that the primary cultures develop and can be maintained for more than 2 weeks. The morphological pattern of these primary cultures is compared to that already described for cells grown in serum-supple mented medium (SSM) [3,10]. In addition, the expression of several neuronal activities present in fetal mouse brain, such as TRH, tyrosine hydroxylase (TH; EC 1.14.16.2), glut amic acid decarboxylase (GAD; EC 4.1.1.15) and choline acety(transferase (ChAT; EC 2.3.1.6) was examined.…”