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bioRxiv · 10.1101/2021.03.31.437811

A new method for obtaining bankable and expandable adult-like microglial cells

You, M.-J.·Rim, C.·Kang, Y.-J.·Kwon, M.
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Abstract

The emerging role of microglia in neurological disorders requires a novel method for obtaining massive amounts of adult microglia because current in vitro methods for microglial study have many limitations, including a limited proliferative capacity, low cell yield, immature form, and too many experimental animals use. Here, we developed a new method for obtaining bankable and expandable adult-like microglial cells using the head neuroepithelial layer (NEL) of mouse E13.5. The NEL includes microglia progenitors that proliferate and ramify over time. Functional validation with a magnetic-activated cell sorting system using the NEL showed that the isolated CD11b-positive cells (NEL-MG) exhibited microglial functions, such as phagocytosis (microbeads, amyloid {beta}, synaptosome), migration, and inflammatory changes following lipopolysaccharide (LPS) stimulation. NEL was subcultured and the NEL-MG exhibited a higher expression of microglia signature genes than the neonatal microglia, a widely used in vitro surrogate. Banking or long-term subculture of NEL did not affect NEL-MG characteristics. Transcriptome analysis revealed that NEL-MG exhibited better conservation of microglia signature genes with a closer fidelity to freshly isolated adult microglia than neonatal microglia. This new method effectively contributes to obtaining adult-like microglial cells, even when only a small number of experimental animals are available, leading to a broad application in neuroscience-associated fields.

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You, M.-J., Rim, C., Kang, Y.-J., Kwon, M.. 2021-03-31. A new method for obtaining bankable and expandable adult-like microglial cells. https://doi.org/10.1101/2021.03.31.437811

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