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Glia toxicity in dissociation cell cultures induced by cyclosporine
G Stoltenburg-Didinger1, F Boegner
1Institute of Neuropathology, Klinikum Steglitz, Free University of Berlin, Germany.
Abstract:
Intravenously applied cyclosporine is the most effective and best analyzed immunosuppressive agent to date. Most frequently, this drug shows nephrotoxic or hepatotoxic side effects, which have already been investigated in detail. In addition to these adverse effects, there is also clear clinical evidence for toxic damage to the central nervous system. On the basis of magnetic resonance tomography and computed tomography studies, the white matter seems to be primarily affected. A systematic approach to neurotoxicity has been established in the following model. Mixed in-vitro cell cultures of dorsal root ganglia (DRG) and of the central nervous system were prepared from 6 to 14 day old chick embryos (E6-E14). For cultivation of nerve and glia cells we used beta NGF, a soluble trophic factor, and NTF B 82 as a matrix factor. Differentiated cultures were incubated with cyclosporine for intravenous application. Within a period of several days up to two weeks the cultures were analyzed using phase contrast microscopy, light microscopy, scanning and transmission electron microscopy. Glia cells and fibroblasts showed the most pronounced toxic effects. Their cytoplasm was infiltrated with smaller and larger vesicles which contained neutral lipids. Control cultures remained unaffected. Because of the close correlation between the in-vitro damage of glia cells and the clinically observed alteration of the white matter, we think our in-vitro model is helpful for the investigation of the neurotoxic effects of cyclosporine and immunotherapeutic drugs.
Insights
Cyclosporine, an immunosuppressant, can cause central nervous system damage. This study developed an in-vitro model using chick embryo nerve and glia cells to investigate cyclosporine
Area of Science:
- Neuroscience
- Immunology
- Toxicology
Background:
- Intravenous cyclosporine is a widely used immunosuppressant with known nephrotoxic and hepatotoxic side effects.
- Clinical evidence suggests cyclosporine can also cause central nervous system toxicity, primarily affecting white matter.
- Previous studies have focused on kidney and liver toxicity, with less investigation into neurotoxicity.
Purpose of the Study:
- To establish and validate an in-vitro model for investigating the neurotoxic effects of cyclosporine.
- To analyze the cellular mechanisms underlying cyclosporine-induced neurotoxicity.
- To provide a reliable model for studying neurotoxicity of immunosuppressive drugs.
Main Methods:
- Preparation of mixed in-vitro cell cultures from chick embryo dorsal root ganglia and central nervous system (E6-E14).
- Cultivation of nerve and glia cells using beta NGF and NTF B 82.
- Incubation of differentiated cultures with cyclosporine and subsequent analysis via microscopy (phase contrast, light, scanning, transmission electron microscopy).
Main Results:
- Glia cells and fibroblasts exhibited the most significant toxic effects.
- Cytoplasmic infiltration with neutral lipid-containing vesicles was observed in affected cells.
- Control cultures without cyclosporine exposure remained unaffected, validating the model's specificity.
Conclusions:
- The developed in-vitro model effectively replicates cyclosporine-induced neurotoxicity observed in clinical settings.
- The model demonstrates that glia cells are primary targets of cyclosporine toxicity.
- This in-vitro system is valuable for further research into the neurotoxic effects of cyclosporine and similar immunotherapeutic agents.