Triple Inhibition of TGFb/BMP/GSK3 Signaling in Human Pluripotent Stem Cells Generates PAX6- Neural Progenitors with robust Self-renewal

In this report, we show that in addition to dual SMAD inhibition, blocking Glycogen synthase kinase 3(GSK3) signaling by adding CHIR99021 generated novel NPCs with significantly enhanced proliferation. By RNA-seq, we show that S/D/C NPCs and S/D NPCs display distinct expression profiles, as S/D/C triggered NPCs are Pax6- and express hindbrain genes while . S/D triggered NPCs are Pax6+ and highly express forebrain genes. Our data reveal that Pax6 labels distinct regionally specified hNPCs and manipulating GSK3 signaling could pattern hNPCs with positionally specified fate and different self-renewal capability. Overall design: We compared the transcriptome of through triple Inhibition of TGFb/BMP/GSK3 Signaling and dual SMAD inhibition derived NPCs by RNA-Seq.