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Myelin pp 131-144 | Cite as

Neuron/Oligodendrocyte Myelination Coculture

  • Yi Pang
  • Kimberly Simpson
  • José Javier Miguel-Hidalgo
  • Renate Savich
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1791)

Abstract

Myelination cell culture systems are useful tools for studying myelin biology and myelin-related disorders. Compared to a number of established protocols for dissociated pure oligodendrocyte (OL) culture, methods for myelination culture are limited. We recently developed a mixed neuron-glia coculture system that generates robust and efficient myelination. By optimizing cell culture conditions, dissociated neural progenitor cells from embryonic rat spinal cords develop into neurons and glial cells including profiles of oligodendrocyte (OL) lineage. Within 4 weeks, OL progenitor cells (OPC) proliferate, differentiate into mature OLs, and myelinate axons. The formation of compact myelin sheath is confirmed by electron microscopy. For morphological analysis by light microscopy, cells grown on glass coverslips are fixed and immunostained for various myelin-related proteins, including those embedded within the myelin sheath and those clustered at the node of Ranvier. Myelinated axons can be quantified readily by either manual counting or ImageJ software. The culture system may also be used for electron microscopic analysis by slightly modifying the cell culture procedure.

Key words

Myelination Oligodendrocyte Axon Embryonic Spinal cord Cell culture 

Notes

Acknowledgments

This work was supported by NIH grants 2R56NS054278, MH084194, and by funds from the Department of Pediatrics, University of Mississippi Medical Center. We thank Glenn Hoskins for his excellent technical support in EM study.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Yi Pang
    • 1
  • Kimberly Simpson
    • 2
    • 3
  • José Javier Miguel-Hidalgo
    • 3
  • Renate Savich
    • 1
  1. 1.Department of PediatricsUniversity of Mississippi Medical CenterJacksonUSA
  2. 2.Department of Neurobiology and Anatomical SciencesUniversity of Mississippi Medical CenterJacksonUSA
  3. 3.Department of Psychiatry and Human BehaviorUniversity of Mississippi Medical CenterJacksonUSA

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