Summary
Cervical and thoracic segments of the spinal cord of six and seven days old chicken embryos were trypsinized. The isolated cells have been cultivated in vitro and studied both light- and electronmicroscopically.
Immediately after isolation, neurons or glial cells can not be distinguished by morphological criteria. After a few hours in vitro, however, the first sprouting of fibres is visible. About 12 hours later an aggregation of cells in bulks or loosely woven nets becomes more and more prominent, without an active cellmovement visible. The aggregational process of the previously dissociated cells is combined with ultrastructural differentiation.
At the end of the first week in vitro one could recognize besides some astrocytes a great number of those cells which were neither astroglia-nor oligodendroglia-cells. The latter were not detectable during our short periods of cultivation.
Those cells showing an enhancement of organelles and their endoplasmic reticulum formed an ergastoplasm after five to seven days in vitro are definitely nerve cells, whose axons and dendrites appear after three days of incubation.
The most convincing criteria of nerve cell maturation is the appearance of synapses, which increase in number from the fifth day of incubation onwards.
Axo-dendritic, axo-somatic and axo-axonal synapses are present.
A possible physiological significance of the synapses in relation to their ultrastructural features and the importance of the “membrane junctions” has been discussed.
Zusammenfassung
Cervikales und thorakales Rückenmark 6 und 7 Tage alter Hühnerembryonen wurde trypsiniert. Die isolierten Rückenmarkszellen wurden kultiviert und anschließend licht- und elektronenmikroskopisch untersucht.
Unmittelbar nach der Isolierung kann man die Zellen morphologisch nicht unterscheiden. Nach wenigen Stunden der Inkubation beobachtet man ein erstes Auswachsen feiner Portsätze. Nach ca. 12stündiger Kulturzeit beginnt eine Zusammenlagerung der Zellen in Zellklumpen und lockeren Zellverbänden immer deutlicher zu werden, ohne daß eine aktive Zellbewegung sichtbar wird. Die Zusammenlagerung der Zellen geht mit einer feinstrukturell verfolgbaren Ausreifung einher.
Am Ende der 1. Woche findet man neben einigen astrocytären Gliazellen auch zahlreiche andere polymorphe Formen, die sich weder als Astroglia- noch als Oligodendroglia-Zellen typisieren lassen. Letztere konnten während der kurzen Inkubationsdauer weder licht- noch elektronenmikroskopisch identifiziert werden.
Die Nervenzellen zeigen eine Zunahme ihrer Organellen und eine Zusammenlagerung ihres endoplasmatischen Retikulums zu Ergastoplasmaformationen. Axone und Dendriten sind bereits nach 3tägiger Inkubation erkennbar.
Ein eindeutiges Kriterium der Nervenzellausreifung stellt die Bildung von Synapsen dar, die nach dem 5. Tag der Inkubation deutlich wird. Es bilden sich axo-dendritische, axosomatische und axo-axonale Synapsen.
Die mögliche physiologische Bedeutung der Synapsen auf Grund ihrer Ultrastruktur und die Bedeutung der ‚'membrane junctions” wird diskutiert.
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Mit Unterstützung durch die Deutsche Forschungsgemeinschaft (Me 276/5).
Die Verff. danken Fräulein R. Dietrich und Frau M. C. Weinrichter für technische Assistenz.
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Meller, K., Breipohl, W., Wagner, H.H. et al. Die Differenzierung isolierter Nerven- und Gliazellen aus trypsiniertem Rückenmark von Hühnerembryonen in Gewebekulturen. Z. Zellforsch. 101, 135–151 (1969). https://doi.org/10.1007/BF00335590
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DOI: https://doi.org/10.1007/BF00335590