Naïve and primed pluripotent stem cells recapitulate the peri- and post-implantation development, respectively. Thus, investigation of distinct traits between each pluripotent stem cell type would shed light on early embryonic processes. Herein, by screening a fluorescent probe library, we found that intracellular glycogen led to specific reactivity to CDg4, a glycogen fluorescence sensor, in both human and mouse naïve embryonic stem cells (ESCs). The requirement of constant inhibition of Gsk3β as well as high oxidative phosphorylation (OxPHOS) in naïve compared to primed ESCs was closely associated to high level of intracellular glycogen in naïve ESCs. Both capacity of OxPHOS and stored glycogen, rescued naïve ESCs by transient inhibition of glycolysis, which selectively eliminated primed ESCs. Additionally, naïve ESCs with active OxPHOS were enriched from a mixture with primed ESCs by high reactivity to ATP-Red1, a mitochondrial ATP fluorescence probe. These results indicate the active OxPHOS and high intracellular glycogen as a novel “biomarker” delineating metabolic remodeling during the transition of naïve pluripotency.

幼稚和已启动的多能干细胞分别概括了植入前后的发育。因此，对每种多能干细胞类型之间不同特征的研究将揭示早期胚胎过程。在此，通过筛选荧光探针库，我们发现细胞内糖原在人和小鼠幼稚胚胎干细胞 (ESC) 中导致对 CDg4（一种糖原荧光传感器）的特异性反应。与致敏 ESC 相比，在幼稚 ESC 中持续抑制 Gsk3β 以及高氧化磷酸化 (OxPHOS) 的要求与幼稚 ESC 中高水平的细胞内糖原密切相关。OxPHOS 的容量和储存的糖原都通过瞬时抑制糖酵解拯救了幼稚的 ESC，从而选择性地消除了已启动的 ESC。此外，通过对线粒体 ATP 荧光探针 ATP-Red1 的高反应性，从与已启动的 ESC 的混合物中富集了具有活性 OxPHOS 的幼稚 ESC。这些结果表明活性 OxPHOS 和高细胞内糖原是一种新的“生物标志物”，描绘了幼稚多能性转变过程中的代谢重塑。