Äussere Körnerzellen des postnatalen Ratten-Cerebellum in vitro : Überleben und Differenzieren in Rein- und Cokulturen
The differentiation and the survival of cerebellar granule cells, co-cultured with either astrocytes or meningeal cells, was monitored by fluorometric measurement of enzyme activities of both, neuron-specific and non-neuronal, enolases as well as glutathione reductase and neuronal nitric oxide synthase. Granule cell viability was monitored using a fluorescein-diacetate assay. Astrocytes are known to stimulate differentiation and survival of cerebellar granule cells, whereas meningeal cells were also being discussed to have a supportive role during cerebellar development. Our results show that diffusible factors from astrocytes induce in co-cultured cerebellar granule cells a significant increase of neuron-specific enolase, and a significant decrease in neuronal nitric oxide synthase activity, whereas glutathione reductase activity was comparable to control cultures without a second population of cells. Granule cell viability was significantly elevated in population co-cultured with astrocytes, which correlates with the low ratio of the activities of nitric oxide synthase and glutathione reductase. Meningeal cells did only exert a moderate change in the respective enzyme activities. Each population of cells used for co-culturing was purified, and separated from the other by a microporous membrane, so that direct cell-to-cell contact was impossible and only diffusible substances could be exchanged between the two cell populations.
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