Figure 9

Sca-1⁺ trophoblast isolated from differentiated TS cell cultures were re-plated in TS media plus growth factors and cultured in 5% CO₂ in air (Normoxia) or 1% O₂ (Hypoxia) for 48 hours. These cultures were compared to proliferating, non-sorted TS cells plated in the same conditions and at the same seeding density (TS). Hypoxia promoted colony formation and proliferation compared to normoxia (A). After 48 hours of culture in hypoxia in the presence of growth factors, expression of Cdx2 and Esrrb in Sca-1⁺ trophoblast was maintained, while Eomes expression was up regulated in these cells as detected by qRT-PCR (B). Cells were stained with hematoxylin and eosin. Scale bar = 500 um.