Metabolic Reprogramming, Autophagy, and Reactive Oxygen Species Are Necessary for Primordial Germ Cell Reprogramming into Pluripotency

Figure 2

(a) EB formation and spontaneous differentiation of reprogrammed PGCs with a low dose of DCA (50 μM) in normoxia. The markers of endoderm, ectoderm, and mesoderm are, respectively, albumin, AE1/AE3 cytokeratins, and vimentin. Scale bars correspond to 25 μm. (b) Confocal microscopy images for immunofluorescence against SSEA1 and HIF1α in PGC cultures. Samples include Cos7 cells cultured in normoxic conditions as a negative control and in hypoxia as a positive control. Images also show PGCs cultured in normoxia (norm), hypoxia (Hyp), and dichloroacetate (50 μM) in normoxia (norm + DCA) for 48 h. Scale bars correspond to 25 μm. (c) Flow cytometry data of PGC cultures showing the percentage of SSEA1⁺ cells displaying green signal from the JC-1 probe (inactive mitochondria), red signal (active mitochondria), or bivalent (both types). An increase in inactive mitochondria in detriment of bivalent mitochondria is observed in normoxia together with either low DCA dose (50 μM), resveratrol (0.5 μM), or VPA (5 mg/mL) and in hypoxia after 5 days with respect to normoxic cultures.

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