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A rapid and reproducible assay for modeling myelination by oligodendrocytes using engineered nanofibers

Nature Protocols volume 8, pages 771–782 (2013)Cite this article

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Abstract

Current methods for studying oligodendrocyte myelination using primary neurons are limited by the time, cost and reproducibility of myelination in vitro. Nanofibers with diameters of >0.4 μm fabricated from electrospinning of liquid polystyrene are suitable scaffolds for concentric membrane wrapping by oligodendrocytes. With the advent of aligned electrospinning technology, nanofibers can be rapidly fabricated, standardized, and configured into various densities and patterns as desired. Notably, the minimally permissive culture environment of fibers provides investigators with an opportunity to explore the autonomous oligodendrocyte cellular processes underlying differentiation and myelination. The simplicity of the system is conducive to monitoring oligodendrocyte proliferation, migration, differentiation and membrane wrapping in the absence of neuronal signals. Here we describe protocols for the fabrication and preparation of nanofibers aligned on glass coverslips for the study of membrane wrapping by rodent oligodendrocytes. The entire protocol can be completed within 2 weeks.

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Figure 1: Fabrication and preparation of the fibers for assaying oligodendroglial membrane wrapping.
Figure 2: Temporal and spatial distribution of OPCs cultured on fibers: proliferation, migration, differentiation and formation of myelin-like segments.

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Acknowledgements

We thank all the members of the Chan laboratory and the Multiple Sclerosis Research Group at the University of California, San Francisco, for encouragement, advice and insightful discussion. This work was supported by the US National Multiple Sclerosis Society Career Transition Award (TA 3008A2/T), the Harry Weaver Neuroscience Scholar Award (JF 2142-A2/T) and a grant from the US National Institute of Health/National Institute of Neurological Disorders and Stroke (NS062796-02) to J.R.C.

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Authors and Affiliations

  1. Department of Neurology and Program in Neuroscience, University of California, San Francisco, San Francisco, California, USA

    Seonok Lee, S Y Christin Chong & Jonah R Chan

  2. Geriatric Research, Education and Clinical Center, Veterans Affairs Ann Arbor Healthcare Center, Ann Arbor, Michigan, USA

    Samuel J Tuck & Joseph M Corey

  3. Department of Biomedical Engineering, University of Michigan, Ann Arbor, Michigan, USA

    Joseph M Corey

  4. Department of Neurology, University of Michigan, Ann Arbor, Michigan, USA

    Joseph M Corey

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  1. Seonok Lee
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  2. S Y Christin Chong
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Contributions

S.L. performed the experiments and analyzed the data. S.J.T. fabricated the fibers. S.L., S.Y.C.C., J.R.C. and J.M.C. provided intellectual contributions and wrote the paper.

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Correspondence to Jonah R Chan.

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The authors declare no competing financial interests.

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Lee, S., Chong, S., Tuck, S. et al. A rapid and reproducible assay for modeling myelination by oligodendrocytes using engineered nanofibers. Nat Protoc 8, 771–782 (2013). https://doi.org/10.1038/nprot.2013.039

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