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

Figure 1

(a) Number of reprogrammed cells in PGC cultures supplemented with DASA (0.1 μM) for 7 days in normoxia. No differences are observed with respect to normoxia. (b) Number of reprogrammed cells in PGC cultures supplemented with DASA (0.1 μM) for 7 days in hypoxia. DASA causes a synergistic effect with hypoxia. (c) Relative expression of PPARγ in PGCs cultured under normoxia or hypoxia. Results are shown normalized with respect to normoxia. (d and e) Number of reprogrammed cells in PGC cultures supplemented with PPARγ agonist ciglitazone (0.1 μM) for 7 days in (d) normoxia or (e) hypoxia. No differences are observed by ciglitazone addition with respect to controls. (f) Number of reprogrammed cells in PGC cultures supplemented with DCA at a high dose (500 μM) for 7 days in normoxia and in hypoxia. DCA prevents hypoxia-induced reprogramming and it has no effect in normoxia. (g and h) Number of reprogrammed cells in PGC cultures supplemented with DCA at a low dose (50 μM) for 7 days. (h) DCA in normoxia is capable of reprogramming, (g) whereas it has no further effect in hypoxia. Asterisks show significance at . (i) Immunofluorescence against phosphorylated PDH (p-PDH) in normoxia (norm) and hypoxia (hyp) with and without 50 μM DCA for 4 days. From left to right: in blue, nuclei stained with DAPI; in red, PGCs detected as SSEA1⁺ cells; in green, p-PDH. Scale bars represent 25 microns.

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