GlutaMAX, 10 ng ml−1 hLIF, 4 μM CHIR99021, 3 μM SB431542 and 0.1 μM Compound E) for another 5 days. The cultures were then split onto Matrigel-coated plates with Accumax (Innovative Cell Technologies) and cultured in Neural Stem cell Maintenance Medium (NSMM) containing 50% Advanced DMEM/F12, 50% Neurobasal, 1 × N2, 1 × B27, 2 mM GlutaMAX, 10 ng ml−1 hLIF, 3 μM CHIR99021 and 2 μM SB431542.
