Bijland, S. et al. (2019) An in vitro model for studying CNS white matter: functional properties and experimental approaches. F1000Research, 8, 117. (doi: 10.12688/f1000research.16802.1) (PMID:31069065) (PMCID:PMC6489523)
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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.
| Item Type: | Articles |
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| Additional Information: | Version 1; peer review: 2 approved. |
| Status: | Published |
| Refereed: | Yes |
| Glasgow Author(s) Enlighten ID: | Muecklisch, Dr Katja and Bijland, Dr Silvia and Crawford, Colin and Anderson, Professor Jim and Barnett, Professor Susan and Linington, Professor Christopher and Thomson, Miss Gemma and McLaughlin, Dr Mark and Edgar, Professor Julia and Euston, Dr Matthew |
| Creator Roles: | Bijland, S.Data curation, Formal analysis, Investigation, Methodology, Project administration, Software, Writing – original draft Thomson, G.Data curation, Formal analysis, Investigation, Methodology, Visualization, Writing – original draft Euston, M.Formal analysis, Methodology, Writing – review and editing Thümmler, K.Investigation, Software Crawford, C. L.Investigation Barnett, S. C.Supervision McLaughlin, M.Funding acquisition, Investigation, Supervision Anderson, T. J.Funding acquisition, Supervision, Writing – review and editing Linington, C.Funding acquisition, Supervision, Writing – review and editing Edgar, J. M.Conceptualization, Data curation, Formal analysis, Funding acquisition, Investigation, Methodology, Project administration, Supervision, Validation, Visualization, Writing – original draft, Writing – review and editing |
| Authors: | 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., and Edgar, J. M. |
| College/School: | College of Medical Veterinary and Life Sciences > School of Infection & Immunity College of Medical Veterinary and Life Sciences > School of Biodiversity, One Health & Veterinary Medicine |
| Research Centre: | College of Medical Veterinary and Life Sciences > School of Infection & Immunity > Centre for Immunobiology |
| Journal Name: | F1000Research |
| Publisher: | F1000Research |
| ISSN: | 2046-1402 |
| ISSN (Online): | 2046-1402 |
| Published Online: | 29 January 2019 |
| Copyright Holders: | Copyright © 2019 Bijland S et al. |
| First Published: | First published in F1000Research 8: 113 |
| Publisher Policy: | Reproduced under a Creative Commons License |
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