Figure 4

TS cell markers, Cdx2 and Esrrb were more highly expressed in Sca-1⁺ trophoblast isolated from differentiated TS cell cultures than in non-sorted, proliferating TS cell cultures (A). Sca-1⁺ trophoblast isolated from differentiated TS cultures gave rise to proliferating trophoblast cell lines that when differentiated by the removal of growth factors, expressed genes indicative of syncytiotrophoblast (Gcm1, Syna), spongiotrophoblast (Tpbpa) and TGCs (Prl3b1) (B). These differentiated cells were also positive for alkaline phosphatase activity (C, blue) and PAS (D, magenta/arrowhead) indicating differentiation of sinusoidal TGCs and glycogen trophoblast, respectively. D0 represents undifferentiated, proliferating TS cells while, D2, D4, D6 represent cultures differentiated for 2, 4 or 6 days. In C, scale bar = 500 um. In D, scale bar = 100 um.