Chunk #23 — ONLINE METHODS — Culture of hiPSCs

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Functional cortical neurons and astrocytes from human pluripotent stem cells in 3D culture.

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hiPSCs were cultured on irradiated DR4 mouse embryonic fibroblast feeders in the following cell culture media: DMEM/F12 (1:1) (Invitrogen) containing 20% KnockOut Serum (Invitrogen), 1 mM non-essential amino acids (Invitrogen, 1:100), GlutaMax (Invitrogen, 1:100), 0.1 nM β-mercaptoethanol (Sigma-Aldrich), 100 U/ml penicillin and 100 μg/ml streptomycin (Invitrogen) and 10–15 ng/ml FGF2 (R&D Systems).