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A culture system to study oligodendrocyte myelination processes using engineered nanofibers

Nature Methods volume 9, pages 917–922 (2012)Cite this article

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Abstract

Current methods for studying central nervous system myelination necessitate permissive axonal substrates conducive to myelin wrapping by oligodendrocytes. We have developed a neuron-free culture system in which electron-spun nanofibers of varying sizes substitute for axons as a substrate for oligodendrocyte myelination, thereby allowing manipulation of the biophysical elements of axonal-oligodendroglial interactions. To investigate axonal regulation of myelination, this system effectively uncouples the role of molecular (inductive) cues from that of biophysical properties of the axon. We use this method to uncover the causation and sufficiency of fiber diameter in the initiation of concentric wrapping by rat oligodendrocytes. We also show that oligodendrocyte precursor cells display sensitivity to the biophysical properties of fiber diameter and initiate membrane ensheathment before differentiation. The use of nanofiber scaffolds will enable screening for potential therapeutic agents that promote oligodendrocyte differentiation and myelination and will also provide valuable insight into the processes involved in remyelination.

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Figure 1: Temporal and spatial behavior of OPCs on polystyrene fibers are similar to that of OPCs on live axons.
Figure 2: Fiber diameter is sufficient to initiate myelination.
Figure 3: Quantification of fiber diameter threshold and size preference for ensheathment and wrapping.
Figure 4: The majority of the cells in the cultures are capable of ensheathing and wrapping fibers above a minimum threshold.

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Acknowledgements

We thank W. Stallcup for the rabbit anti-PDGFRα antibody; M.L. Wong for sectioning nanofiber cultures for electron microscopy analysis at the W.M. Keck Foundation Advanced Microscopy Laboratory at UCSF; R. Langen and M. Isas for assistance, advice and support with the electron microscopy; the other members of the Chan laboratory and the MS Research Group at UCSF for encouragement, advice and insightful discussions. 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 the US National Institutes 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, University of California, San Francisco (UCSF), San Francisco, California, USA

    Seonok Lee, Stephanie A Redmond, S Y Christin Chong & Jonah R Chan

  2. Program in Neuroscience, UCSF, San Francisco, California, USA

    Seonok Lee, Stephanie A Redmond, S Y Christin Chong & Jonah R Chan

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

    Michelle K Leach, Zhang-Qi Feng & Joseph M Corey

  4. Department of Obstetrics, Gynecology and Reproductive Sciences, UCSF, San Francisco, California, USA

    Synthia H Mellon

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

    Samuel J Tuck & Joseph M Corey

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

    Joseph M Corey

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  1. Seonok Lee
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Contributions

S.L., M.K.L., S.A.R., S.Y.C.C. and J.R.C. performed experiments. S.L., M.K.L., S.H.M., S.J.T., Z.-Q.F., J.M.C. and J.R.C. provided reagents. S.L., M.K.L., S.A.R., S.Y.C.C., S.H.M., S.J.T., Z.-Q.F., J.M.C. and J.R.C. provided intellectual contributions. S.L. and J.R.C. analyzed the data and wrote the paper.

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Correspondence to Joseph M Corey or Jonah R Chan.

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The authors declare no competing financial interests.

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Lee, S., Leach, M., Redmond, S. et al. A culture system to study oligodendrocyte myelination processes using engineered nanofibers. Nat Methods 9, 917–922 (2012). https://doi.org/10.1038/nmeth.2105

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