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

Martina von der Bey; Silvia De Cicco; Susanne Zach; Bastian Hengerer; Ebru Ercan-Herbst

doi:10.1016/j.xpro.2023.102164

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

STAR Protoc. 2023 Mar 17;4(2):102164. doi: 10.1016/j.xpro.2023.102164. Online ahead of print.

Authors

Martina von der Bey¹, Silvia De Cicco², Susanne Zach³, Bastian Hengerer³, Ebru Ercan-Herbst⁴

Affiliations

¹ BioMed X Institute, Im Neuenheimer Feld 515, 69120 Heidelberg, Germany; Molecular and Translational Neuroscience, Department of Neurology, Ulm University, N27, Albert-Einstein-Allee 11, 89081 Ulm, Germany.
² BioMed X Institute, Im Neuenheimer Feld 515, 69120 Heidelberg, Germany.
³ Boehringer Ingelheim Pharma GmbH & Co. KG, Birkendorfer Str. 65, 88397 Biberach an der Riss, Germany.
⁴ BioMed X Institute, Im Neuenheimer Feld 515, 69120 Heidelberg, Germany. Electronic address: ercan@bio.mx.

Abstract

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.

Keywords: Cell Biology; Cell Differentiation; Cell culture; Cell isolation; Microscopy; Neuroscience; Stem Cells.