High yield primary microglial cultures using granulocyte macrophage-colony stimulating factor from embryonic murine cerebral cortical tissue

Journal of Neuroimmunology
Adam C YuJacqueline A Quandt

Abstract

Microglia play vital roles in neurotrophic support and modulating immune or inflammatory responses to pathogens or damage/stressors during disease. This study describes the ability to establish large numbers of microglia from embryonic tissues with the addition of granulocyte-macrophage stimulating factor (GM-CSF) and characterizes their similarities to adult microglia examined ex vivo as well as their responses to inflammatory mediators. Microglia were seeded from a primary embryonic mixed cortical suspension with the addition of GM-CSF. Microglial expression of CD45, CD11b, CD11c, MHC class I and II, CD40, CD80, and CD86 was analyzed by flow cytometry and compared to those isolated using different culture methods and to the BV-2 cell line. GM-CSF microglia immunoreactivity and cytokine production was examined in response to lipopolysaccharide (LPS) and interferon-γ (IFN-γ). Our results demonstrate GM-CSF addition during microglial culture yields higher cell numbers with greater purity than conventionally cultured primary microglia. We found that the expression of immune markers by GM-CSF microglia more closely resemble adult microglia than other methods or an immortalized BV-2 cell line. Primary differences amongst the differ...Continue Reading

Citations

Dec 17, 2019·NPJ Parkinson's Disease·Sandeep K BarodiaMatthew S Goldberg
Jan 10, 2020·Naunyn-Schmiedeberg's Archives of Pharmacology·Bin HuiQinhua Zhou
May 21, 2020·International Journal of Molecular Sciences·Dorothea R MorrisJiyang Cai
Dec 11, 2020·NPJ Parkinson's Disease·Sandeep K BarodiaMatthew S Goldberg
May 1, 2018·Journal of Neuroscience Methods·Marieta GeorgievaWenlong Huang
Dec 3, 2021·Glia·Hadar LeviAri Barzilai

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