Myelin pp 179-192 | Cite as

A Central Nervous System Axonal Myelination Assay for High-Throughput Screening

  • Karen Lariosa-WillinghamEmail author
  • Dmitri Leonoudakis
Part of the Methods in Molecular Biology book series (MIMB, volume 1791)


The formation of new myelin in persistent multiple sclerosis (MS) lesions is compromised, leading to a reduction in neuron function and subject to degeneration and death. Current MS therapies can control autoimmune-mediated demyelination, but none directly promote the regeneration of myelin in the central nervous system (CNS). To identify new drugs that stimulate remyelination, we established a high-throughput cell-based assay to identify compounds that promote myelination. Methods were developed for initiating myelination in vitro using a preparation of primary embryonic rat cortical cells. We developed an immunofluorescent phenotypic image analysis method to quantify the morphological alignment of myelin characteristic of the initiation of myelination. The assay scalability and consistency was validated by screening the NIH clinical collection library of 727 compounds and identified ten compounds that promote myelination (Lariosa-Willingham et al., BMC Neurosci 17:16, 2016). Here, we present the detailed methods for a high capacity in vitro assay that assesses myelination of live axons.

Key words

Myelination Oligodendrocyte High throughput Drug screening Differentiation Primary cell-based assay Image analysis Multiple sclerosis Myelin basic protein 


  1. 1.
    Deshmukh VA, Tardif V, Lyssiotis CA, Green CC, Kerman B, Kim HJ, Padmanabhan K, Swoboda JG, Ahmad I, Kondo T, Gage FH, Theofilopoulos AN, Lawson BR, Schultz PG, Lairson LL (2013) A regenerative approach to the treatment of multiple sclerosis. Nature 502(7471):327–332. Scholar
  2. 2.
    Mei F, Fancy SPJ, Shen YA, Niu J, Zhao C, Presley B, Miao E, Lee S, Mayoral SR, Redmond SA, Etxeberria A, Xiao L, Franklin RJM, Green A, Hauser SL, Chan JR (2014) Micropillar arrays as a high-throughput screening platform for therapeutics in multiple sclerosis. Nat Med 20(8):954–960. Scholar
  3. 3.
    Najm FJ, Madhavan M, Zaremba A, Shick E, Karl RT, Factor DC, Miller TE, Nevin ZS, Kantor C, Sargent A, Quick KL, Schlatzer DM, Tang H, Papoian R, Brimacombe KR, Shen M, Boxer MB, Jadhav A, Robinson AP, Podojil JR, Miller SD, Miller RH, Tesar PJ (2015) Drug-based modulation of endogenous stem cells promotes functional remyelination in vivo. Nature 522(7555):216–220. Scholar
  4. 4.
    Banker G, Goslin K (1998) Culturing nerve cells. Cellular and molecular neuroscience, 2nd edn. MIT Press, CambridgeGoogle Scholar
  5. 5.
    Demerens C, Stankoff B, Logak M, Anglade P, Allinquant B, Couraud F, Zalc B, Lubetzki C (1996) Induction of myelination in the central nervous system by electrical activity. Proc Natl Acad Sci U S A 93(18):9887–9892CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Fedoroff S, Richardson A (2001) Protocols for neural cell culture, 3rd edn. Humana Press, Totowa, NJCrossRefGoogle Scholar
  7. 7.
    Lubetzki C, Demerens C, Anglade P, Villarroya H, Frankfurter A, Lee VM, Zalc B (1993) Even in culture, oligodendrocytes myelinate solely axons. Proc Natl Acad Sci U S A 90(14):6820–6824CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Watkins TA, Emery B, Mulinyawe S, Barres BA (2008) Distinct stages of myelination regulated by gamma-secretase and astrocytes in a rapidly myelinating CNS coculture system. Neuron 60(4):555–569. Scholar
  9. 9.
    Lariosa-Willingham KD, Rosler ES, Tung JS, Dugas JC, Collins TL, Leonoudakis D (2016) Development of a central nervous system axonal myelination assay for high throughput screening. BMC Neurosci 17:16. Scholar

Copyright information

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

Authors and Affiliations

  1. 1.Teva Pharmaceuticals Biologics DiscoveryRedwood CityUSA

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