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On DIV 3, the medium was replaced with Neurobasal medium (Thermo Fisher Scientific) supplemented with B-27 (Thermo Fisher Scientific), glutaMAX (Thermo Fisher Scientific), primocin (0.1 g/ml, Invivogen), doxycycline (4 g/ml, Sigma Aldrich), BDNF (10 ng/ml, Bioconnect), NT3 (10 ng/ml, Bioconnect) and forskolin (10 M, Sigma Aldrich)

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