Fig 7. Neural priming and CHIR99021 treatment efficiently induce differentiation of P/G-iPSCs into GFP-positive cells.

A, Schematic diagram of the culture procedure for differentiation. Undifferentiated P/G-iPSCs cultured for 3 days in N2/B27 medium in the presence or absence of 1 μM RA and/or 1 mM VPA were dissociated into single cells in 0.25% trypsin-EDTA and quickly reaggregated (3000 cells/150 μl/well) using 96-well low-adhesion plates in GMEM supplemented with 10% KSR in the presence of 10 μM CHIR99021. The aggregates were incubated for 10 days at 37°C with 5% CO₂. B–E, Representative image of GFP-expression. Scale bars indicate 100 μm. F, After 10 days of culture with CHIR99021, the percentages of GFP-positive cells were determined. **P<0.01. *P<0.05. n.s.: Difference not significant (P>0.05). Values represent the mean ± S.E. from five independent experiments. CHIR, CHIR99021. PD, PD0325901. RA, retinoic acid. VPA, valproic acid.