Cuprizone-Induced Demyelination and Remyelination Models of Multiple Sclerosis

Multiple sclerosis (MS) is marked by central nervous system inflammation, demyelination, and neuronal injury, leading to cumulative neurological deficits. Despite reducing relapse rates, current therapies have limited impact on slowing disability accumulation in progressive forms of MS and do not directly promote remyelination. The cuprizone model is one of the most widely used demyelination models for studying oligodendrocyte death, myelin loss, and spontaneous remyelination. Remyelination is often impaired in MS lesions, and because cuprizone-induced demyelination often recovers quickly, it does not model inflammation-mediated inhibition of remyelination. Therefore, cuprizone can be combined with adoptive transfer of activated myelin-specific CD4⁺ T cells at the onset of spontaneous repair, and this combination model is referred to as the adoptive transfer-cuprizone (ATCup) paradigm, which recapitulates the immune-mediated suppression of remyelination seen in MS. This chapter provides detailed protocols for the cuprizone and ATCup paradigms, including immune cell generation and transfer, tissue processing, and quantitative histology. These models enable mechanistic studies of demyelination and repair and facilitate testing of drugs that protect oligodendrocytes and promote remyelination.