Generation of a Definitive Endoderm enriched population from mouse ES cells.

<p>(<b>A</b>) Protocol for generation of definitive endoderm (DE) in mouse embryonic stem (mES) cells. Cells were cultured as embryoid bodies in chemically defined medium and treated for 2 days with 100 ng/ml Activin A (A) and 10 ng/ml BMP4 (B), and for the following 2 or 4 days with Activin A alone. (<b>B</b>) Treated (B+A) and non treated (N/A) cells were harvested at days 2, 4 and 6 of the differentiation protocol and expression of definitive endoderm (Sox17 and Foxa2), extraembryonic endoderm (Sox7), mesendoderm (Mixl1 and Gsc) and mesoderm (Bry T) markers was determined by RT-qPCR. Data represent the average of triplicate experiments and are expressed as mean ± SEM. Data are expressed as gene expression relative to Gapdh and normalised against day 0 mouse embryonic stem cells (mESCs), where p<0.05 (*), p<0.01(**) or p<0.001 (***).