The neural progenitor cell proliferation and differentiation cultures were generated from telencephalic tissue derived from embryonic day (E) 15–17. Pregnant dams, either alcohol drinking (prenatal alcohol exposure; PAE) or saccharin drinking (control) were anesthetized via inhalation of isofluorane in a small chamber, and using a midline transverse abdominal incision, the gravid uterus was removed and rinsed in chilled 1X DPBS (no calcium, no magnesium; Gibco; 14190136). All embryos from the same dam were isolated and rinsed in chilled 1X DPBS solution, and the number of embryos and crown-to-rump length (mm) were recorded. Whole brains were removed and placed into chilled 1X Hank's Balanced Salt Solution (HBSS) with glucose and magnesium chloride (Cellgro; 21-023-CV). Olfactory bulbs and meninges were removed, and the telencephalons were collected into a sterile conical tube filled with cold HBSS. After complete collection, HBSS was aspirated and pre-warmed medium was added. Medium consisted of Dulbecco's Modified Eagle's Medium with Nutrient Mixture (Ham's) F-12 medium (DMEM/F12) with glutamine (Invitrogen; 11320-082) supplemented with N2 (Invitrogen; 17502-048), 20 ng/mL FGF-2 (Peprotech; 100-18B), 20 ng/mL EGF (Peprotech; AF-100-15), and 1% antibiotic-antimycotic
