Neuron-oligodendrocyte myelination co-culture derived from embryonic rat spinal cord and cerebral cortex
Article first published online: 30 JAN 2012
© 2012 The Authors. Brain and Behavior published by Blackwell Publishing Ltd.
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Brain and Behavior
Volume 2, Issue 1, pages 53–67, January 2012
How to Cite
Pang, Y., Zheng, B., Kimberly, S. L., Cai, Z., Rhodes, P. G. and Lin, R. C. S. (2012), Neuron-oligodendrocyte myelination co-culture derived from embryonic rat spinal cord and cerebral cortex. Brain and Behavior, 2: 53–67. doi: 10.1002/brb3.33
- Issue published online: 30 JAN 2012
- Article first published online: 30 JAN 2012
- Received: 6 October 2011; Revised: 13 December 2011; Accepted: 18 December 2011
An in vitro myelination model derived from rat central nervous system (CNS) remains to be established. Here, we describe a simple and reproducible myelination culture method using dissociated neuron-oligodendrocyte (OL) co-cultures from either the embryonic day 16 (E16) rat spinal cord or cerebral cortex. The dissociated cells are plated directly on poly-L-lysine-coated cover slips and maintained in a modified myelination medium that supports both OL and neuron differentiation. The spinal cord derived OL progenitor cells develop quickly into myelin basic protein (MBP)+ mature OLs and start to myelinate axons around 17 days in vitro (DIV17). Myelination reaches its peak around six weeks (DIV40) and the typical nodes of Ranvier are revealed by paranodal proteins Caspr and juxaparanodal protein Kv1.2 immunoreactivity. Electron microscopy (EM) shows typical myelination cytoarchitecture and synaptic organization. In contrast, the cortical-derived co-culture requires triiodothyronine (T3) in the culture medium for myelination. Finally, either hypomyelination and/or demyelination can be induced by exposing proinflammatory cytokines or demyelinating agents to the co-culture, suggesting the feasibility of this modified in vitro myelination model for myelin-deficit investigation.