4.8. Cell Culture

Both neuronal and mixed glial neuronal cultures were prepared from embryonic mice at 13–14 days as described previously [53]. In brief, the growth medium consisted of Dulbecco’s Modified Eagle Medium (DMEM, GibcoBRL, Grand Island, NY, USA) with 2 mM glutamine, 5% fetal bovine serum, and 5% horse serum. Minced cerebral cortices combined with growth medium were seeded onto a poly-D-lysine (Sigma, St. Louis, MO, USA) pre-coated plate at 8–9 hemispheres per 24-well plate. The cultures were incubated at 37 °C in a humidified 5% CO₂ atmosphere. All cultures were used at 10–14 days in vitro. These experiments were performed under the guidelines for the care and use of mice in research and under protocols approved by the Animal Care and Use Committee of Sejong University.