BV2 Microglial Cell Line Cultures

The immortalized mouse microglial cell line (BV2) was originally obtained from the Cell Resource Centre, Peking Union Medical College. BV2 cells were cultured in DMEM/high glucose. 24 h after plated at a density of 35,000 cells/ml, BV2 cells were primed with different doses of mouse recombinant BAFF (“BAFF priming”) for 24 h. Cells were pre-incubated for at least an hour with 1 mM 2-Deoxy-d-glucose (2-DG) or 100 nM rapamycin before priming to block glycolysis or mTOR, respectively. The medium was changed after 24 h and cells were rested for 5 days. On day 6, cells were re-stimulated with 100 ng/ml LPS (“LPS restimulation”) or 40 ng/ml BAFF (“BAFF restimulation”). All the compounds were dissolved in 0.1% DMSO in DMEM/high glucose because 2-DG and rapamycin were dissolved in DMSO. The control group is 0.1% DMSO in DMEM/high glucose and described as “CTL.” The cells were incubated in a 5% CO2 incubator maintained at 37°C. 24 h after the last stimulation, cellular supernatants or cell lysates were harvested.