Investigating demyelination, efficient remyelination and remyelination failure in organotypic cerebellar slice cultures: Workflow and practical tips

Rianne P. Gorter*, Naomi S. Dijksman, Wia Baron, Holly Colognato

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapterAcademicpeer-review


Healthy myelin is essential for proper brain function. When the myelin sheath is damaged, fast saltatory impulse conduction is lost and neuronal axons become vulnerable to degeneration. Thus, regeneration of the myelin sheath by encouraging oligodendrocyte lineage cells to remyelinate the denuded axons is a promising therapeutic target for demyelinating diseases such as multiple sclerosis. Ex vivo organotypic cerebellar slice cultures are a useful model to study developmental myelination, demyelination, remyelination and remyelination failure. In these cultures, the cerebellum's three-dimensional architecture and various cell populations remain largely intact, providing a realistic and relatively cost-efficient model that can be easily manipulated by the addition of viral vectors, pharmaceuticals or (transgenic) cells to augment or replace resident cell populations. Moreover, slice cultures can be treated with lysolecithin or polyinosinic:polycytidylic acid to induce demyelination and mimic efficient as well as inefficient remyelination. It can be challenging to set up slice cultures for the first time, as in our experience, seemingly minor differences in technique and materials can make a great difference to the quality of the cultures. Therefore, this report provides an in-depth description for the generation and maintenance of ex vivo organotypic cerebellar cultures for demyelination-remyelination studies with a focus on practical tips for scientists that are new to this technique.

Original languageEnglish
Title of host publicationMethods in Cell Biology
EditorsDavid C. Montrose
Number of pages21
ISBN (Electronic)9780323899468
ISBN (Print)9780323899451
Publication statusPublished - 30-Mar-2022

Publication series

NameMethods in Cell Biology
ISSN (Print)0091-679X


  • Cerebellar slice cultures
  • Demyelination
  • Ex vivo model
  • Lysolecithin
  • Multiple sclerosis
  • Myelin
  • Myelination
  • Organotypic slice cultures
  • Remyelination
  • Rodent

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