Российская академия наук


Cerebellum organotypic cultivation by the method of Victorov. An attempt of multiple sclerosis modeling in culture



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Cerebellum organotypic cultivation by the method of Victorov. An attempt of multiple sclerosis modeling in culture

Fetisova E.K., Avetisyan A.V. Samoilova D.V., Kireev R.I.

Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow, Russia, ekfetisova@gmail.com


Damage of oligodedrocytes (CNS producer of myelin) may lead to demyelination and –as a result - to neurodegenerative diseases, the most common of which is a multiple sclerosis (MS). Demylienation is considered to be a central event in MS pathology. The cellular mechanisms of demylianation in RS are largely unknown. In vitro cultivation of nervous tissue is a simplified way to study of de – and re- myelination processes, caused by pathological and pharmacological action. It is well known that the complex of interactions between neurons, oligodendrocytes and – possibly astrocytes - are needed for myelin formation. Therefore only few alternative in vitro models are available for de- and re - myelination study. Neuronal and glial monolayer cultures are not available. Organotypic cultures are seemed to be the most adequate ones. Cell organization and interactions in such cultures are closely related to in vivo ones. Cerebellum organotypic cultivation of new born rats by the method of Victorov was performed. Cerebellum (1mmx1mm) pieces were seeded on the collagen covered glasses. Cultural medium was supplied with chicken embryonic extract and fetal bovine sera. It allows avoiding growth factor addition. In the course of in vitro cultivation the cells are totally deprived of monoaminergic effects, which are present in natural conditions. In agreement with Chubakov works, the cultural medium was supplied with serotonin to stimulate myelination. Fibroblast growth factor, known to cause axon growth, was added beginning with14-16 cultiation day. Oligodedrocytes were identified by special “Oligo” marker. Myelination level was tested by incubation with antibodies to Myelin Basic protein at 21 – 28 day of cultivation.

Lipopolysaccharid (LPS) was used as a specific inducer of demyelination. LPS-model might represent the type III oligodedropathology that is seen in a subset of MS patients. Nitric oxid synthase formation after LPS treatment was tested by staining with specific antibodies.



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