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A Step-by-step Protocol for Obtaining Mature Microglia from Mice.

Min-Jung You1, Min-Soo Kwon1

  • 1Department of Pharmacology, Research Institute for Basic Medical Science, School of Medicine, CHA University, CHA BIO COMPLEX, 335 Pangyo, Bundang-gu, Seongnam-si, Gyeonggi-do 13488, Republic of Korea.

Bio-Protocol
|September 9, 2022
PubMed
Summary

Researchers developed a novel method to culture adult-like microglia (NEL-MG) from mouse neuroepithelial layers. This technique provides higher yields than existing in vitro models for studying brain immune cells.

Keywords:
BankingExpandable cellsMicrogliaMicroglial progenitorNeuroepithelial layer (NEL)

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Area of Science:

  • Developmental neuroscience
  • Immunology
  • Stem cell biology

Background:

  • Microglia, the brain's resident immune cells, originate from yolk sac precursors.
  • These precursors migrate to the brain and differentiate within the neuroepithelial layer (NEL) during embryonic development.
  • Existing in vitro models for microglia research have limitations in yield and maturity.

Purpose of the Study:

  • To establish a novel in vitro system for generating mature microglia.
  • To utilize the neuroepithelial layer (NEL) as a source for microglial precursors.
  • To compare the yield and characteristics of derived microglia with existing models.

Main Methods:

  • Dissection of the embryonic mouse neuroepithelial layer (NEL) at E13.5.
  • Culture and expansion of NEL cells to promote microglial precursor maturation.
  • Isolation of mature microglia (NEL-MG) using magnetic-activated cell sorting (MACS) for CD11b-positive cells.

Main Results:

  • The NEL was found to be bankable and expandable, supporting cell culture.
  • Microglial precursors within the NEL matured with extended culture duration.
  • Higher yields of adult-like microglia (NEL-MG) were obtained compared to neonatal microglia and cell lines.

Conclusions:

  • The neuroepithelial layer (NEL) serves as a viable source for generating mature microglia in vitro.
  • NEL-derived microglia (NEL-MG) represent a promising in vitro model for studying microglial function.
  • This method offers an improved yield of adult-like microglia for research applications.