Cited by 85Neuroinflammation and Neurodegeneration MechanismsPluripotent Stem Cells ResearchNeurogenesis and neuroplasticity mechanisms

Generation of microglial cells from mouse embryonic stem cells

Beutner, Clara, Roy, Kristin, Linnartz, Bettina, Napoli, Isabella, Neumann, Harald·2010·Nature Protocols

Abstract

Microglia, the resident immune cells of the brain, are difficult to obtain in high numbers and purity using currently available methods; to date, microglia for experimental research are mainly isolated from the brain or from mixed glial cultures. In this paper, we describe a basic protocol for the in vitro differentiation of mouse embryonic stem (ES) cells into microglial precursor cells. Microglia are obtained by a protocol consisting of five stages: (i) cultivation of ES cells, (ii) formation and differentiation of embryoid bodies, (iii) differentiation into neuroectodermal lineage and isolation of myeloid precursor cells, (iv) differentiation into microglial precursor cells and (v) cultivation of ES cell-derived microglial precursors (ESdMs). The protocol can be completed in 60 d and results in stably proliferating ESdM lines, which show inducible transcription of inflammatory genes and cell marker expression comparable with primary microglia. Furthermore, ESdMs are capable of chemokine-directed migration and phagocytosis, which are major functional features of microglia.

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Crossref · 10.1038/nprot.2010.90

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  • These genes are expressed at low levels in cultured macrophage cell lines and previously described mouse ES-derived microglia.

    “Notably, these genes are expressed at low levels in cultured macrophage cell lines as well as in previously described mouse ES-derived microglia 33.”

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Authors
Beutner, C., Roy, K., Linnartz, B., Napoli, I., & Neumann, H.
Affiliations
University of Bonn (DE)
Corresponding author
Neumann, Harald
Venue
Nature Protocols
Published
2010-08-05
Volume
5
Issue
9
Pages
1481-1494
ISSN
  • 1750-2799 (ISSN-L)
  • 1754-2189
OpenAlex
W2044415255
Cited by
85
References
18
Citations by year
2026: 1 · 2024: 1 · 2023: 2 · 2022: 3 · 2021: 5 · 2020: 5 · 2019: 3 · 2018: 8 · 2017: 10 · 2016: 8 · 2015: 10 · 2014: 8 · 2013: 10 · 2012: 6
FWCI
4.06
Type
journal article
Language
en
Open access
closed
Locations
  • Nature Protocols (publishedVersion)
  • PubMed (publishedVersion)
Topics
  • Life Sciences › Neuroscience › Neurology › Neuroinflammation and Neurodegeneration Mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
  • Life Sciences › Neuroscience › Developmental Neuroscience › Neurogenesis and neuroplasticity mechanisms
Keywords
microglia, mouse embryonic stem cells, in vitro differentiation, embryoid bodies, neuroectodermal differentiation, myeloid precursors, phagocytosis, inflammatory gene expression
MeSH terms
Animals, Cell Differentiation, Culture Media, Mice, Inbred C57BL, Microglia, Cell Culture Techniques, Mice, Embryonic Stem Cells

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