A novel cell line from spontaneously immortalized murine microglia

作者：Nagamoto Combs Kumi^*; Kulas Joshua; Combs Colin K

来源：Journal of Neuroscience Methods, 2014, 233: 187-198.

DOI：10.1016/j.jneumeth.2014.05.021

• Microglia
• Inflammation
• Lipopolysaccharide
• Beta-amyloid
• Phagocytosis
• M1/M2 phenotype

Background: Purified microglia cultures are useful tools to study microglial behavior in vitro. Microglial cell lines serve as an attractive alternative to primary microglia culture, circumventing the costly and lengthy preparation of the latter. However, immortalization by genetic or pharmacologic manipulations may show altered physiology from primary microglia. %26lt;br%26gt;New method: A novel microglial cell line was isolated from a primary glial culture of postnatal murine cerebral cortices. The culture contained a population of spontaneously transformed microglia that continued to divide without genetic or pharmacological manipulations. After several clones were isolated, one particular clone, SIM-A9, was analyzed for its microglial characteristics. %26lt;br%26gt;Results: SIM-A9 cells expressed macrophage/microglia-specific proteins, CD68 and Iba1. SIM-A9 cells were responsive to exogenous inflammatory stimulation with lipopolysaccharide and beta-amyloid, triggering tyrosine kinase-based and NF kappa B signaling cascades as well as TNF alpha. secretion. SIM-A9 cells also exhibited phagocytic uptake of fluorescent labeled beta-amyloid and bacterial bioparticles. Furthermore, lipopolysaccharide increased the levels of inducible nitric oxide synthase and cyclooxygenase-2, whereas IL-4 stimulation increased arginase-1 levels demonstrating that SIM-A9 cells are capable of switching their profiles to pro- or anti-inflammatory phenotypes, respectively. %26lt;br%26gt;Comparison with existing methods: The use of SIM-A9 cells avoids expensive and lengthy procedures required for the preparation of primary microglia. Spontaneously immortalized SIM-A9 cells are expected to behave more comparably to primary microglia than virally transformed or pharmacologically induced microglial cell lines. %26lt;br%26gt;Conclusions: SIM-A9 cells exhibit key characteristics of cultured primary microglia and may serve as a valuable model system for the investigation of microglial behavior in vitro.

• 出版日期2014-8-15