engleski

Morphological and functional changes in BV-2 microglia due to various stimuli

Microglia cells represent the innate immunity in the central nervous system. They are reactive by nature, and as such are affected by all changes in their environment. Various stimuli can activate microglia cells through pattern recognition receptors. They can be divided into two groups: pathogen-associated molecular patterns (PAMP) and danger-associated molecular patterns (DAMP). In-vitro cellular environments are unable to imitate the one present in the central nervous system, which leads to morphological and functional changes to the microglial cells. Murine microglial BV-2 cell line is commonly used as a substitute for primary microglia. They are cultivated in the Dulbecco’s modified eagle medium (DMEM) supplemented with fetal calf serum (FCS). Aim of our study was to determine how various stimuli alter the morphology and function of BV-2 microglial cells. The stimuli used for this research were DAMP and PAMP signals: murine cytomegalovirus (MCMV) as a PAMP signal, while DAMPs were fetuin A, as a component of FCS, and HSP70, as a marker of cellular stress in hypoxia. Cells were cultivated in varying FCS percentage (0%, 5% and 10%), while also being infected with MCMV. Results from our study show that manipulation of cultivation conditions, specifically FCS depletion, leads to morphological and functional changes of BV-2 microglia cells. Complete FCS depletion resulted in a resting microglial state, while also increasing susceptibility of this cell line for MCMV-infection. Varying FCS depletion also affected the levels of pro-inflammatory markers CD86 and iNOS, depending on the concentration. Hypoxic cells displayed a predominant inflammatory profile in the early post-hypoxia period, with a tendency to an anti-inflammatory switch in the late post-hypoxia periods. This indicates that the environmental changes that modulate the metabolic and immunological profile, which can cause an altered response of microglial cells to pathogens.

microglia ; morphology ; DAMP / PAMP stimuli

nije evidentirano