Human fetal neuron cultures and neurospheres (hNSPs) were obtained from the Basic Science Core I of the Comprehensive NeuroAIDS Center (CNAC)– headed by Dr. Kamel Khalili at Temple University Lewis Katz School of Medicine. hNSPs were cultured and maintained in DMEM/F12 media supplemented with N2 supplement, FGF, Glutamax, pen/strep, and Hepes (5mM). hNSPs were expanded and cryopreserved for future expansion and usage. Neural progenitors (NPCs) were cultured from hNSPs as follows: hNSPs were dissociated by trypsin, filtered through cell strainers (100 mic), and plated on Geltrex coated dishes in NPC differentiation media consist of neurobasal media supplemented with B27, FGF, glutamax, Na-pyruvate, N-acetyl-L-cysteine, and pen/strep. hNPCs cultures were expanded and cryopreserved for future expansion and usage. hNPCs and hNSPs were characterized by immunocytochemical analysis of GFAP, Nestin, and Sox2 staining. Immature and mature neuron cultures were prepared directly from the matching fetal brains by differentiation. Fetal human brain cells were plated in culture dishes coated with laminin and Poly-L-ornitine. Cells were differentiated into neurons by neuronal differentiation media consist of neurobasal media supplemented with B27, glutamax, BDNF, GDNF, and pen/strep
