Three-dimensional co-culture platform of human induced pluripotent stem cell-derived oligodendrocyte lineage cells and neurons for studying myelination

Developing an in vitro platform to study neuron-oligodendrocyte interaction, particularly myelination, is essential to understand aberrant myelination in neuropsychiatric and neurodegenerative diseases. Here, we provide a controlled, direct co-culture protocol for human induced-pluripotent-stem-cell (hiPSC)-derived neurons and oligodendrocytes on three-dimensional (3D) nanomatrix plates. We describe steps to differentiate hiPSCs into cortical neurons and oligodendrocyte lineage cells on 3D nanofibers. We then detail the detachment and isolation of the oligodendrocyte lineage cells, followed by neuron-oligodendrocyte co-culture in this 3D microenvironment.