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

Figure 5

Effect of antibiotic treatment on the expression of markers of definitive endoderm and hepatic progenitor fate. H9 cells (passage number 39) were differentiated towards definitive endoderm and hepatic fate in the presence of antibiotics throughout the course of differentiation as indicated. (a) Cells were collected for RNA isolation on day 5 (endoderm) and day 9 (hepatic progenitors) of differentiation in the presence of antibiotics and qRT-PCR was performed to analyze the expression of endoderm specific markers, Foxa2 and Sox17, and hepatic progenitor markers, HNF4α and GATA4, using gene specific primers. The expression of Polr2g gene was used as normalizer. The bars represent normalized fold mRNA expression with values of untreated samples set as 1. The results are representative of three independent differentiation experiments. (b) The protein expression levels of Foxa2 and Sox17 (samples from day 5) and HNF4α (samples from day 9) were analyzed using Western blot for samples treated with antibiotic concentrations as above. The expression of GAPDH was used as loading control. The results are representative of three independent experiments.

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