Differentiation of Neural Cell Lines

ReN VM neural progenitor (catalog number SCC008) and Neuro-2a mouse neuroblast (catalog number CCL-131) cells were obtained from Millipore and ATCC (VA, USA), respectively. The undifferentiated ReN VM cells were grown on Matrigel basement membrane matrix (catalog number 354230, Corning, NY, USA) and maintained in DMEM/F12 (catalog number 11320033, Thermo Fisher Scientific) supplemented with 20ng/ml basic fibroblast growth factor (catalog number 8910, Cell Signaling, MA, USA), 20ng/ml epidermal growth factor (catalog number RKP01133, Reprokine, FL, USA), 10 Units/ml heparin sodium salt (catalog number H3149, Sigma-Aldrich) and 1X N21-MAX (catalog number AR008, R&D Systems, MN, USA) or 1X B27 media supplement (catalog number 17504044, Thermo Fisher Scientific). The cells were differentiated into neuronal and glial populations with the removal of heparin and growth factors from media for at least 7 days. Passage-matched N2a and RML-infected N2a cells were created as described previously [64 (link)] and maintained in DMEM (catalog number 11995065, Thermo Fisher Scientific), supplemented with 10% fetal bovine serum (catalog number 12483020, Thermo Fisher Scientific) and 1% GlutaMAX (catalog number 35050061, Thermo Fisher Scientific). CRISPR-Cas9-generated N2a knockout cells were described previously [65 (link)].