Metabolic activity affects RPE fate during cell reprogramming

To investigate transcriptomic changes that facilitate the reprogramming of retinal pigment epithelium (RPE) cells to neural retina, chicken RPE explants were cultured for 24 or 48 hours in the presence or absence of the reprogramming factor FGF2. To further interrogate the metabolic requirements of reprogramming, RPE explants were additionally cultured in the presence of the pyruvate dehydrogenase kinase inhibitor dichloroacetate in the presence or absence of FGF2. Overall design: RPE explants were collected from specific pathogen free chickens at embryonic day 4. Biological replicates were collected in triplicate. Three explants were pooled per biological replicate. Explants were treated with A) maintenance media (control), B) 10 ng / ml FGF2, c) 20 mM dichloroacetate, or D) both 10 ng / ml FGF2 and 20 mM dichloroacetate. Each condition was assayed at both 24 and 48 hours in culture.