The dynamic differentiation of hES cells to DE is influenced by ECMPs.

<p>(<b>a)</b> Schematic overview of the semi-optimal protocol for DE differentiation. hES cells were seeded and kept undifferentiated for 4 days to allow expansion. Subsequently the cells were treated with Wnt3a to direct the cells from the pluripotent stage into the mesendoderm stage, followed by 3 days with Activin A to direct the cells from the mesendoderm stage to the DE stage. <b>(b)</b> Representative immunofluorescence images from three independent experiments of cells cultured on the five different ECMP combinations at different time points during the DE differentiation protocol. The tested ECMP combinations were fibronectin (Fn), collagen II+fibronectin (Col2+Fn), collagen I (Col1), netrin 1+fibronectin (Ne+Fn) and vitronectin (Vn) (scale bar = 200μm). <b>(c)</b> Magnification of cultures on Col1 at day 5 (scale bar = 200μm).