Ex Vivo Slice Cultures to Study Myelination, Demyelination, and Remyelination in Mouse Brain and Spinal Cord

In vitro culture systems have been invaluable in understanding the cell biology of oligodendrocytes; the monoculture of primary oligodendroglia has helped characterize different stages of oligodendrocyte maturation in the absence of neurons. However, oligodendrocyte monocultures do not model the interaction of oligodendrocytes with neurons where they form myelin wraps. To circumvent this problem, coculture systems were developed; oligodendrocytes and neurons are cultured together, facilitating the study of myelin wraps and the interaction between the two cell types. However, this coculture system also has limitations, as other cells are not present and it does not represent the three-dimensional multicellular structure seen in vivo. Some of these limitations are resolved by using ex vivo slice cultures to serve as a three-dimensional culture system that is more similar to in vivo and can be used to study myelination, demyelination, and remyelination, over extended periods of time. Slice cultures are economical compared to in vivo studies and live imaging using them is less challenging. The focus of this chapter is to describe how to culture brain and spinal cord slices of mice and use them to study myelination, demyelination, and remyelination.