DMEM/F-12

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Generation of iPSC-derived cortical rosettes: Differentiation of iPSC-derived cortical rosettes was performed as described with slight adaptations (Shi et al., 2012). In brief, iPSC were cultured in PluriPro medium (Cell Guidance Systems). Once the cell culture reached 95% confluence, neural induction was initiated by changing the culture medium to neural induction media containing DMEM/F12 (N2 supplement; 1:50), Neurobasal (B27 supplement; 1:50) mixed at a 1:1 ratio and cAMP (300 ng/ml, Sigma-Aldrich), LDN-193189 (0.5 μM, Miltenyi), A83-01 (0.21 μg, Miltenyi) and XAV939 (2 μM, Enzo). Cells were maintained in this medium for 8–11 days, collected by dissociation with TrypL Express (Invitrogen) and replated in neural differentiation media containing DMEM/F12 (N2 supplement; 1:50), Neurobasal (B27 supplement; 1:50) mixed at a 1:1 ratio and cAMP (300 ng/ml, Sigma Aldrich) on Geltrex-coated (GT, Life Technologies) plastic dishes. For analyzing neurogenesis rosettes were passaged every 5 days in a 1:3 ratio. Quantification of ßIII-tubulin positive cells was performed at day 2 of each passage. For blocking of N-cadherin function cells were plated at 140.000/cm2 on GT coated plastic dishes in neural differentiation media containing 20 μg/ml N-cadherin blocking Antibody (Sigma, C3865), 25 μg/ml BDNF and 12.5 μg/ml GDNF (both Cell Guidance Systems). N-cadherin activation was performed by plating cells at 140.000/cm2 on plastic dishes coated with GT and 5 μg/ml N-cadherin protein (1388-NC, R&D Systems). Half of the media was changed on a daily basis. Quantification of ßIII-tubulin positive cells was performed at day 6.

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