Implementation of a toxic cuprizone model of demyelination in vitro

Home/2018, Vol. 6, No. 1/Implementation of a toxic cuprizone model of demyelination in vitro

Cell and Organ Transplantology. 2018; 6(1):93-98.
DOI: 10.22494/COT.V6I1.84

Implementation of a toxic cuprizone model of demyelination in vitro

Rodnichenko A. E.
State Institute of Genetic and Regenerative Medicine of the National Academy of Medical Sciences of Ukraine, Kyiv, Ukraine

The relevance of the use of adequate experimental models to study the mechanisms of the development of multiple sclerosis and the effects of drugs on the remyelination of nerve fibers due to the prevalence of this disease, which affects mostly young people.
The purpose of the study was to adjustment a toxic cuprizone model in vitro, with confirming the processes of demyelination of nerve fibres.
Materials and methods. To study the features of the processes of myelination and demyelination of neurons axons, the culture of dissociated cerebellar cells of 7-day old newborn FVB/N mice was used. To detect myelin sheaths, we used a histochemical staining with Sudan Black B dye. To identify oligodendrocytes, their culture was stained by immunocytochemistry for Olig2 marker.
Results. Developing a model of toxic demyelination, we determined the density of the cerebellar cell culture, selected conditions for the cultivation of dissociated cerebellar cells, and the survival of the dissociated culture of the cerebellum until the 26-28th day of cultivation was achieved. Using histochemical staining, the presence of myelination and demyelination processes in vitro was confirmed. Using immunocytochemical staining, it has been shown that the neurotoxin cuprizone results in a decrease of Olig2-positive oligodendrocytes, when it is applied on the 18th day of cultivation of the dissociated cerebellar cells culture.
Conclusion. The conditions for the cultivation of murine dissociated cerebellar cells were selected, the presence of myelination and demyelination processes in vitro was confirmed; a decrease in the number of Olig2-positive cells when the demyelination agent was added to the culture on the 18th day was shown.

Key words:  cuprizone; cerebellum; oligodendrocytes; demyelination

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Rodnichenko A. Implementation of a toxic cuprizone model of demyelination in vitro. Cell and Organ Transplantology. 2018; 6(1):93-98. doi:10.22494/cot.v6i1.84

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