Generating Neural Progenitor Cells from hESCs

To generate NPCs, human embryonic stem cells BR-1 (Fraga et al., 2011 (link)) (kindly provided by Prof. Lygia Pereira, São Paulo University - USP) were grown on polystyrene plates (TPP, Switzerland) covered with Matrigel (BD Biosciences, Franklin Lakes, NJ, USA) in StemPro medium containing 8 ng/mL basic fibroblast growth factor and 0.1 mM β-mercaptoethanol (all from Thermo Fischer Scientific, Waltham, MA, USA). After propagation, the cells were differentiated as neural cells using inhibitors of bone morphogenetic protein (Noggin; R&D Systems, USA), and transforming growth factor-beta (SB431542; Tocris Bioscience, Bristol, UK) (Chambers et al., 2009 (link)). At this moment, cells showed morphology and expression of markers consistent to those of neural progenitor cells Figs. S2A and S2B.