Mouse glia were cultured from the cortex of newborn CD1 mice. The cortex was dissected, digested with papain 10 U/ml for 20 minutes at 37°C, and harshly triturated. Cells were plated onto T75 flasks in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS). After reaching 80–90% confluence, glial cultures were trypsinized and re-plated at lower density (1:3–4) in DMEM + 10% FBS. This re-plating procedure was repeated 2–3 times (typically 3–4 days apart) to remove mouse neurons before cultured glia were used for analysis or co-culture experiments with human neurons. No antibiotics or other drugs were used in glial cultures.
