Figure 1: Generation and neural differentiation of DS iPSCs.

(a) A schematic procedure for directed and spontaneous differentiation of DS iPSCs to neurons and astroglia. Insets (from left to right): representatives bright-field images showing the embryoid bodies (EBs), neural rosette, neurosphere and differentiated neurons, and astroglia under directed and spontaneous differentiation conditions. Scale bars, 200 μm. (b,c) Representatives of iPSCs derived from DS patients expressing pluripotent markers Oct4 and SSEA4, as well as Nanog and Tra-1-81. (d) Representative of DS iPSC-derived NPCs expressing Pax6 and Nestin. (e,f) Representatives of βIII-tubulin+ neurons and A2B5+ glial progenitors derived from DS NPCs under directed differentiation conditions. (g) Representatives of astroglia differentiated under directed astroglial differentiation condition from control (Cont) and DS iPSCs expressing CD44 and vimentin, as well as S100B and GFAP. (h) Representatives of βIII-tubulin+ neurons and S100B+ astroglia derived from DS and Cont NPCs under spontaneous differentiation conditions. Scale bars, 50 μm. (i,j) Quantification of pooled data from Cont and DS lines showing the percentage of βIII-tubulin+ neurons and S100B+ astroglia derived from DS and Cont NPCs (n=3–5 from each cell line), and the length of the longest neurites of neurons (n=10 from each cell line) under spontaneous differentiation conditions. Student’s t-test, *P<0.05 and **P<0.01. Blue, 4′,6-diamidino-2-phenylindole dihydrochloride (DAPI)-stained nuclei. Data are presented as mean±s.e.m.