Figure 2

hES cell-derived precursors express neural markers and differentiate into mature neurons in vitro . (A) Neural precursors at day 11 after the beginning of SMAD inhibition; 96% ± 3% of cells expressed nestin (n = 4), 75% ± 7% expressed PAX6 (n = 4), and 64% ± 9% expressed SOX1 (n = 3). Scale bars are 30 μm. (B) After re-plating for terminal differentiation, cells positive for NeuN (red) and NF (green) were evident after 7 days (a) and persisted through 28 days (b). These cells formed large networks across the culture, and co-localization of NeuN and NF was evident (c). Scale bars are 10 μm. (C) At 28 days, cells expressed the AMPA and NMDA receptors GluA1, GluN1, GluN2B, and GluN3A. GluA1 and GluN2B are shown co-localized with NeuN and NF, respectively. Nestin was still present in these cultures through 21 days. Scale bars are 10 μm.