Figure 2.

Analysis of hPSC-Derived Neural Crest

(A and B) Putative neural crest derived from H9:SOX10 and sorted for GFP expression (A) upregulates CD73, CD90, and CD105 in mesenchymal growth conditions (see Experimental Procedures) and (B) can be differentiated into osteocytes and chondrocytes, stained with Alizarin red and toluidine blue, respectively.

(C and D) H9:SOX10-derived neural crest in neural differentiation conditions are positive for peripherin and TUJ-1 in unsorted cells via intermediate spheroid culture (C) and in sorted cells alongside sensory neural markers BRN3a and ISL1 (D).

(E) Expression of the glial markers SOX10 and S100b in neural crest derived from the iPSC line NB1.

(F) Expression of the cranial neural crest marker ETS-1 at day 7 of neural crest induction (Miff-1).

(G) In vitro, putative neural crest are highly motile; scratch ablation recolonized by GFP positive cells (arrow) within 16 hr (H9:SOX10).

(H) Sections of typical H9:SOX10-derived neural crest spheres after 4 weeks of non-adherent culture.