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. Author manuscript; available in PMC: 2013 Oct 21.
Published in final edited form as: Cell Stem Cell. 2012 Jun 28;11(2):264–278. doi: 10.1016/j.stem.2012.04.027

Figure 1. HD Fibroblasts are Reprogrammed into Karyotypically Normal iPSCs that Generate NSCs.

Figure 1

(A) All reprogrammed lines form colonies (brightfield) and express the pluripotency markers Oct4, Tra-1-60 and SSEA4 by immunocytochemistry (ICC). (B) G-banding showed that the HD180i.5 line had a normal karyotype, which was representative of all lines at the colony and NSC stages. (C) Schematic of the different differentiation protocols used. (D) CAG repeat analysis in HD109i.1 fibroblasts and NSCs over 26 passages showed a small increase in repeat length over time. (E) Western blots of HTT expression in iPSC-derived NSCs with the HTT antibody 2166 demonstrate normal (bottom arrow) and mutant (top arrow) HTT (epitope: amino acids 441–455). (F) Western blots of polyglutamine expressionin iPSC-derived NSCs using the IC2 antibody demonstrates mutant HTT with expanded repeats in the HD-derived lines. (G) Representative image of a NSC sphere demonstrating the section sampled for (H). (H) HD iPSC-derived NSCs can be expanded as spherical aggregates in a self-renewing condition. ICC on cryosections of NSC aggregates demonstrated a consistent expression of the neural progenitor markers PAX6 and nestin in the three lines. (I) Hierarchical clustering of top 1601 genes from NSCs is represented by the vertical bars (yellow for HD and green for control). The dataset shows that HD and control NSC lines are separated into two clusters, confirming the differential expression of these genes into the two categories.