4.3.1. Cell Culture

BV-2 cells were used for in vitro cell experiments. We cultured the cells with a culture medium of Dulbecco s Modified Eagle Medium (DMEM) (Gibco, Grand Island, NY, USA), supplemented with 2.2 g sodium bicarbonate, 4.8 g HEPES (4-(2-hydroxyethyl)-1piperazineethanesulfonic acid) buffer adjusted at pH 7.2–7.4, antibiotic-antimycotic 100× (Biowest, France), L-glutamine, and 10% (*v*/*v*) of fetal bovine serum (FBS) (Gibco, Grand Island, USA). The BV-2 cells were cultured in 6-well plates at the density of 10<sup>5</sup> cells/cm2 and incubated under 5% CO2 at 37 ◦C. The activation was induced by lipopolysaccharide (LPS).

The shape of BV-2 cells was round and irregular spindle when they were not stimulated and stimulated by LPS.
