Human ES Cell Culture Conditions Fail to Preserve the Mouse Epiblast State

Figure 2

Human ESC/EpiSC culture condition matures the ESCs further to a primitive streak-like state. (a) Expression levels of the specific pluripotency-associated genes in the indicated samples. AA/F2 treatment led to the induction of primed pluripotency genes and downregulation of PGC and naïve pluripotency genes. (b) Analysis of pluripotency genes in AA/F2-treated cells in the indicated samples. Gapdh and Pmm2: housekeeping genes. Rex1, Sox2, and Nanog were downregulated, and Oct4 expression was retained. Error bars: (); , , , and (, , and ) (Student’s -test). (c) Heat map showing the expression status of lineage-specific markers in the indicated samples in comparison with naïve mESCs. PS markers are induced in AA/F2-3 days (3D). (d) Analysis of EpiSCs (Fgf5) and PS markers (T and Fgf8) in the indicated samples. Fgf5 and Fgf8 were induced right from 1D, and by 6D, the PS marker T was also highly induced. Error bars: (); , , , and ( and ) (Student’s -test). (e) Western blot analysis of the indicated markers in the indicated samples. Six days of AA/F2 treatment led to high protein expression of the PS/mesendoderm markers T and Eomes and downregulation of Sox2. (f) Representative immunostainings of the indicated proteins in the indicated samples (scale bars 100 μm).

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