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Trophoblast differentiation

AA Aybuke Alici-Garipcan
BÖ Burcu Özçimen
IS Ilke Süder
VÜ Volkan Ülker
TÖ Tamer T. Önder
NÖ Nesrin Özören
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iPS cells were maintained routinely on MEFs with hES media (10 ng/mL FGF2). For the trophoblast differentiation, 2.4 × 104 cells per square centimeter were seeded on Matrigel coated plates with conditioned hES medium by a monolayer of mitomycin-C treated MEF feeder cells (MEF-CM) containing FGF2 (10 ng/mL). On the next day, medium was changed to MEF-CM containing 4 ng/mL of FGF2. Next day, the medium was changed to BMP4 (10 ng/mL; RD Systems), the ALK4/5/7 inhibitor, A83-01 (1 μM; Tocris), and the FGF2-signaling inhibitor PD173074 (0.1 μM; Sigma) containing (BAP) hESC basal medium not conditioned with MEF feeder cells22,27,28. Control cultures were grown in the presence of FGF2 and in the absence of BAP. The medium was replenished daily.

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