Neuroblastoma Cells and Astrocytes Protocol

Mouse brain neuroblastoma cells (N2-A) and rat primary astrocytes (DI-TNC1) were purchased from American Type Culture Collection (ATCC, Manassas, VA). N2- A cell line used in the current investigation is a neuronal cell line known for its high lactate production compare to other cell lines. We, as well as others, have used this cell line and is considered an appropriate model to evaluate potential anti-cancer agents [22 (link), 23 (link)]. We also used the N2-A cell line to investigate the “Warburg Effect” phenomenon [24 (link)], and cancer cells metabolism [25 (link), 26 (link)]. On the other hand, the DI-TNC1 is an astrocyte immortal cell line with lower lactate efflux production compared to N2-A cells, an observation in our lab. The DI-TNC1 is very important in controlling brain energy metabolism [27 (link), 28 (link)]. Cell culture Dulbecco's Modified Eagle Medium (DMEM), fetal bovine serum (FBS), penicillin/streptomycin, DPBS, and trypsin were all from Atlanta Biologicals (Atlanta, GA, USA). Cells were cultured in 75-cm TC flask at 37°C in humidified 5% CO₂ incubator and were subcultured as needed with trypsin/EDTA. Growing media was supplemented with 10% FBS (v/v), 4 mM L-glutamine, and 1% penicillin /streptomycin.