Effects of Aminoglycoside Antibiotics on Human Embryonic Stem Cell Viability during Differentiation In Vitro

Figure 2

Establishment of directed differentiation of H9 cells towards neural fate using dual SMAD inhibition protocol. (a) Cells were collected from day 0 (passage number 37) and day 12 (passage number 39) of differentiation for immunostaining using antibodies against Oct4 and Pax6. The cells were imaged with fluorescence microscope. DAPI represents nuclear staining. Scale bar = 100 μm. (b) Cells were collected for RNA isolation from days 0, 7, and 12 of differentiation. The mRNA expression of pluripotency markers, Oct4 and Nanog, and neural stem cell-specific markers Pax6, Emx2, Pou3f2, and Otx2 were analyzed by qRT-PCR using gene-specific primers. The bars represent fold expression of indicated genes over normalizing control, Polr2g. Values are expressed as mean ± SD in each group. The results are representative of at least three independent differentiation experiments.

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