The next stage of complexity from the OPC HCS Screening Assay incorporates the use of CNS slices, which have the benefit of 3D CNS architecture and cellular environment. The CNS Slice model has a valuable place for the investigation of compounds effect on remyelination efficiency as a translational link between in-vitro cell culture and in-vivo investigations.
Cerebellar slices from neonatal mice undergo spontaneous myelination in ex vivo slice cultures. Lysophosphatidyl choline (LPC) is added to the cultures to demyelinate axons. Remyelination efficiency after treatment with test compounds can be evaluated using immunohistochemistry (Myelin Basic Protein (MBP) to label myelin and neurofilament (NF) to label axons) to quantify the amount of myelin per axon. Confocal microscopy and automated image analysis are used to generate the ‘remyelination index’ for each treatment group.