Generation of microglial cells from mouse embryonic stem cells
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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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.”
From Efficient derivation of microglia-like cells from human pluripotent stem cells (2016) · cites this paper as [33]
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Rett Syndrome Induced Pluripotent Stem Cell-derived Neurons Exhibit Electrophysiological Aberrations
Farra, N. (2012) · TSpace (University of Toronto)
Establishment of polarized microglial cells derived from mouse embryonic stem cells
Ruegsegger, C. (2010) · Infoscience (Ecole Polytechnique Fédérale de Lausanne)
Investigation of the endocannabinoid system using in vivo and in vitro models
Ternes, S. (2013) · bonndoc (University of Bonn)
Roy, K. (2013) · bonndoc (University of Bonn)
Self-citationOligodendrocytes as active modulators of neuroinflammation
Janssen, K., & Beyer, C. (2014) · RWTH Publications (RWTH Aachen)
Optimization of a neuron-microglia co-culture model to explore cell-to-cell interaction dynamics
Vogiatzis, S., Severa, M., Pietrantoni, A., Cairo, G., Falco, R. P., Veroni, C., Lanciotti, A., Etna, M. P., Abate, D. A., Nencioni, L., et al. (2026) · Frontiers in Immunology · doi:10.3389/fimmu.2026.1850795
- Authors
- Beutner, C., Roy, K., Linnartz, B., Napoli, I., & Neumann, H.
- Affiliations
- University of Bonn (DE)
- Corresponding author
- Neumann, Harald
- ORCID
- 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
- Licence
- 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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