Fig. 12.

Immunocytochemical analysis of the otocyst-like cells induced from H9 hESCs. (A) Schematic figure of the culture conditions used to differentiate prosensory cells from induced otic placodal/otocyst cells. The cells were grown in culture with 25 ng/mL FGF3, 25 ng/mL FGF50 from Day 7 to Day 22, and 200 nM RA from Day 7 to Day 14. Three different culture conditions were compared (20 nM RA, 200 nM RA or no RA treatment from Day 14 to Day 22). (B) Representative immunocytochemical images of induced cells on Day 25 of culture. The cells were coimmunostained for the otic placodal markers SIX1, PAX2 and SOX2. The nuclei were stained with Hoechst 33,342. Scale bars, 50 μm. (C) Representative immunocytochemical images of induced cells on Day 25 of culture. The cells were coimmunostained for the otic placodal markers EPCAM, JAG1 and SOX2. The nuclei were stained with Hoechst 33,342. Scale bars, 50 μm.