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Method Article
Martin Frasch
University of Washington
Corresponding Author
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Neuroinflammation in utero may result in life-long neurological disabilities. Microglia play a pivotal role, but the mechanisms are poorly understood. No early postnatal treatment strategies exist to enhance neuroprotective potential of microglia. An important step toward that goal is establishing adequate pathophysiological models where microglia function in a large, well gyrated mammalian brain can be studied. Here we present the protocol to manipulate ovine fetal microglia in primary cultures in vitro without or with previous exposure to endotoxin (lipopolyssacharide, LPS) in vivo. We present the approach of blocking or stimulating the α7 nicotinic acetylcholine receptor (α7nAChR) in fetal microglia which allows to enhance or reduce their pro-inflammatory phenotype when incubated with LPS. The measurement of cytokine IL-1beta in supernatant and the RNAseq pipeline are presented.
Keywords
CHRNA7, microglia, fetus, sheep, RNAseq, inflammation
Figure 1
None to declare.
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Associated Publications
α7 nicotinic acetylcholine receptor signaling modulates the inflammatory phenotype of fetal brain microglia: first evidence of interference by iron homeostasis
by M. Cortes, M. Cao, H. L. Liu, +8
Scientific Reports (28 September, 2017)
RNAseq profiling of primary microglia and astrocyte cultures in near-term ovine fetus: A glial in vivo-in vitro multi-hit paradigm in large mammalian brain
by M. Cortes, M. Cao, H.L. Liu, +7
Journal Of Neuroscience Methods (28 September, 2017)
Fetal microglial phenotype in vitro carries memory of prior in vivo exposure to inflammation
by Mingju Cao, Marina Cortes, Craig S. Moore, +9
Method Article
Martin Frasch
University of Washington
Corresponding Author
Version History
CURRENT STATUS: Posted
Version 1
Posted 01 Feb, 2018
No community comments so far
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Neuroinflammation in utero may result in life-long neurological disabilities. Microglia play a pivotal role, but the mechanisms are poorly understood. No early postnatal treatment strategies exist to enhance neuroprotective potential of microglia. An important step toward that goal is establishing adequate pathophysiological models where microglia function in a large, well gyrated mammalian brain can be studied. Here we present the protocol to manipulate ovine fetal microglia in primary cultures in vitro without or with previous exposure to endotoxin (lipopolyssacharide, LPS) in vivo. We present the approach of blocking or stimulating the α7 nicotinic acetylcholine receptor (α7nAChR) in fetal microglia which allows to enhance or reduce their pro-inflammatory phenotype when incubated with LPS. The measurement of cytokine IL-1beta in supernatant and the RNAseq pipeline are presented.
Figure 1
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Associated Publications
α7 nicotinic acetylcholine receptor signaling modulates the inflammatory phenotype of fetal brain microglia: first evidence of interference by iron homeostasis
by M. Cortes, M. Cao, H. L. Liu, +8
Scientific Reports (28 September, 2017)
RNAseq profiling of primary microglia and astrocyte cultures in near-term ovine fetus: A glial in vivo-in vitro multi-hit paradigm in large mammalian brain
by M. Cortes, M. Cao, H.L. Liu, +7
Journal Of Neuroscience Methods (28 September, 2017)
Fetal microglial phenotype in vitro carries memory of prior in vivo exposure to inflammation
by Mingju Cao, Marina Cortes, Craig S. Moore, +9