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

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
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 Anderson, Professor Jim and Barnett, Professor Susan and Linington, Professor Christopher and McLaughlin, Dr Mark and Edgar, Dr Julia and Crawford, Colin and Thomson, Miss Gemma 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 > Institute of Infection Immunity and Inflammation
College of Medical Veterinary and Life Sciences > School of Veterinary Medicine
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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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
465821The development of an in vitro assay to test the consequences of the acute ablation of myelin proteins on axonal integrity.Julia EdgarMultiple Sclerosis Society (MS)853/07III -IMMUNOLOGY
513131An investigation of the role of microglia/macrophages in axonal damage, in the context of demyelinationJulia EdgarMultiple Sclerosis Society (MS)918/09III -IMMUNOLOGY
633391A high throughput to identify small moleculesJulia EdgarMultiple Sclerosis Society (MS)991III -IMMUNOLOGY