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

• Published in: STAR protocols 2023-03, Vol.4 (2), p.102164-102164, Article 102164
• Main Authors: von der Bey, Martina, De Cicco, Silvia, Zach, Susanne, Hengerer, Bastian, Ercan-Herbst, Ebru
• Format: Article
• Language: English
• Subjects: Cell Biology; Cell culture; Cell Differentiation; Cell isolation; Microscopy; Neuroscience; Protocol; Stem Cells
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2023.102164

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. [Display omitted] •Generation of hiPSC-derived cortical neurons on 3D nanofiber network•Generation of hiPSC-derived oligodendrocyte lineage cells on 3D nanofiber network•Detachment and isolation of hiPSC-derived oligodendrocyte lineage cells•Co-culture of hiPSC-derived oligodendrocyte lineage cells and neurons in 3D environment Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. 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.