In:
Frontiers in Cellular Neuroscience, Frontiers Media SA, Vol. 14 ( 2020-12-21)
Abstract:
Microglia are central nervous system (CNS) resident immune cells that have been implicated in neuroinflammatory pathogenesis of a variety of neurological conditions. Their manifold context-dependent contributions to neuroinflammation are only beginning to be elucidated, which can be attributed in part to the challenges of studying microglia in vivo and the lack of tractable in vitro systems to study microglia function. Organotypic brain slice cultures offer a tissue-relevant context that enables the study of CNS resident cells and the analysis of brain slice microglial phenotypes has provided important insights, in particular into neuroprotective functions. Here we use RNA sequencing, direct digital quantification of gene expression with nCounter® technology and targeted analysis of individual microglial signature genes, to characterize brain slice microglia relative to acutely-isolated counterparts and 2-dimensional (2D) primary microglia cultures, a widely used in vitro surrogate. Analysis using single cell and population-based methods found brain slice microglia exhibited better preservation of canonical microglia markers and overall gene expression with stronger fidelity to acutely-isolated adult microglia, relative to in vitro cells. We characterized the dynamic phenotypic changes of brain slice microglia over time, after plating in culture. Mechanical damage associated with slice preparation prompted an initial period of inflammation, which resolved over time. Based on flow cytometry and gene expression profiling we identified the 2-week timepoint as optimal for investigation of microglia responses to exogenously-applied stimuli as exemplified by treatment-induced neuroinflammatory changes observed in microglia following LPS, TNF and GM-CSF addition to the culture medium. Altogether these findings indicate that brain slice cultures provide an experimental system superior to in vitro culture of microglia as a surrogate to investigate microglia functions, and the impact of soluble factors and cellular context on their physiology.
Type of Medium:
Online Resource
ISSN:
1662-5102
DOI:
10.3389/fncel.2020.592005
DOI:
10.3389/fncel.2020.592005.s001
DOI:
10.3389/fncel.2020.592005.s002
DOI:
10.3389/fncel.2020.592005.s003
DOI:
10.3389/fncel.2020.592005.s004
DOI:
10.3389/fncel.2020.592005.s005
DOI:
10.3389/fncel.2020.592005.s006
DOI:
10.3389/fncel.2020.592005.s007
DOI:
10.3389/fncel.2020.592005.s008
DOI:
10.3389/fncel.2020.592005.s009
Language:
Unknown
Publisher:
Frontiers Media SA
Publication Date:
2020
detail.hit.zdb_id:
2452963-1
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