4.7 Article

A new method for obtaining bankable and expandable adult-like microglia in mice

Journal

JOURNAL OF NEUROINFLAMMATION
Volume 18, Issue 1, Pages -

Publisher

BMC
DOI: 10.1186/s12974-021-02351-4

Keywords

Microglia; Neuroepithelial layer; Subculture; Banking

Funding

  1. National Research Foundation of Korea (NRF) - Korea government (MIST) [2019M3C7A1032561]
  2. Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI) - Ministry of Health & Welfare, Republic of Korea [HI16C1559]
  3. Basic Science Research Program [2020R1F1A1074668]
  4. National Research Foundation of Korea [2019M3C7A1032561] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)

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This study developed a new method to obtain mature microglia from mouse NEL at E13.5, which exhibited microglial functions and maintained characteristics in passage culture. Transcriptome analysis showed NEL-MG had better conservation of microglia signature genes compared to neonatal microglia.
Background The emerging role of microglia in neurological disorders requires a novel method for obtaining massive amounts of adult microglia. We aim to develop a new method for obtaining bankable and expandable adult-like microglia in mice. Methods The head neuroepithelial layer (NEL) that composed of microglial progenitor and neuroepithelial cells at mouse E13.5 was dissected and then cultured or banked. Microglia (MG) isolated from the cultured NEL by magnetic-activated cell sorting system were obtained and named NEL-MG. Results The NEL included microglia progenitors that proliferate and ramify over time with neuroepithelial cells as feeder. In functional analysis, NEL-MG exhibited microglial functions, such as phagocytosis (microbeads, amyloid beta, synaptosome), migration, and inflammatory response following lipopolysaccharide (LPS) stimulation. NEL was passage cultured 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 passage culture 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. NEL-MG could be re-expandable when they were plated again on neuroepithelial cells. Conclusions This new method effectively contributes to obtaining sufficient matured form of microglia (adult-like microglia), even when only a small number of experimental animals are available, leading to a broad application in the field of neuroscience.

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