An in vitro model for studying CNS white matter: functional properties and experimental approaches

Silvia Bijland, Gemma Thomson, Matthew Euston, Kyriakos Michail, Katja Thümmler, Steve Mücklisch, Colin L. Crawford, Susan C. Barnett, Mark McLaughlin, T. James Anderson, Christopher Linington, Euan R. Brown, Eric R. Kalkman, Julia M. Edgar

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2 Citations (Scopus)
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Abstract

The normal development and maintenance of CNS white matter, and its responses to disease and injury, are defined by synergies between axons, oligodendrocytes, astrocytes and microglia, and further influenced by peripheral components such as the gut microbiome and the endocrine and immune systems. Consequently, mechanistic insights, therapeutic approaches and safety tests rely ultimately on in vivo models and clinical trials. However, in vitro models that replicate the cellular complexity of the CNS can inform these approaches, reducing costs and minimising the use of human material or experimental animals; in line with the principles of the 3Rs. Using electrophysiology, pharmacology, time-lapse imaging, and immunological assays, we demonstrate that murine spinal cord-derived myelinating cell cultures recapitulate spinal-like electrical activity and innate CNS immune functions, including responses to disease-relevant myelin debris and pathogen associated molecular patterns (PAMPs). Further, we show they are (i) amenable to siRNA making them suitable for testing gene-silencing strategies; (ii) can be established on microelectrode arrays (MEAs) for electrophysiological studies; and (iii) are compatible with multi-well microplate formats for semi-high throughput screens, maximising information output whilst further reducing animal use. We provide protocols for each of these. Together, these advances increase the utility of this in vitro tool for studying normal and pathological development and function of white matter, and for screening therapeutic molecules or gene targets for diseases such as multiple sclerosis, motor neuron disease or spinal cord injury, whilst avoiding in vivo approaches on experimental animals.
Original languageEnglish
Article number117
JournalF1000Research
Volume8
DOIs
Publication statusPublished - 29 Jan 2019

Keywords

  • (re)myelination
  • microelectrode array
  • microglia
  • semi-high throughput
  • siRNA
  • spinal cord

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)
  • Pharmacology, Toxicology and Pharmaceutics(all)

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    Bijland, S., Thomson, G., Euston, M., Michail, K., Thümmler, K., Mücklisch, S., Crawford, C. L., Barnett, S. C., McLaughlin, M., Anderson, T. J., Linington, C., Brown, E. R., Kalkman, E. R., & Edgar, J. M. (2019). An in vitro model for studying CNS white matter: functional properties and experimental approaches. F1000Research , 8, [117]. https://doi.org/10.12688/f1000research.16802.1