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

Figure 4

Establishment of directed differentiation of H9 cells towards hepatic fate. H9 cells (passage number 37) were differentiated towards hepatic fate using published protocols in feeder free condition on Matrigel-coated dishes. (a) Cells were collected from day 0, day 5, and day 9 of differentiation for immunostaining using antibodies against pluripotency marker Oct4; endoderm specific markers, Foxa2 and Sox17; and hepatic progenitor marker, HNF4α. The cells were imaged with fluorescence microscope. DAPI represents nuclear staining. Scale bar = 100 μm. (b) The mRNA expression of pluripotency markers, Oct4 and Sox2; and definitive endoderm-specific markers, Foxa2 and Sox17; and hepatic progenitor markers, HNF4α; and GATA4 were analyzed by qRT-PCR using gene specific primers. The bars represent fold expression of indicated genes over normalizing control, Polr2g. Error bars represent standard deviation. The results are representative of at least three independent differentiation experiments.

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