One of the most important properties of human embryonic stem cells (hESCs) is related to their primed and naive pluripotent states. Our previous meta-analysis indicates the existence of heterogeneous pluripotent states derived from diverse naive protocols. In this study, we have characterized a commercial medium (RSeT)-based pluripotent state under various growth conditions. Notably, RSeT hESCs can circumvent hypoxic growth conditions as required by naive hESCs, in which some RSeT cells (e.g., H1 cells) exhibit much lower single cell plating efficiency, having altered or much retarded cell growth under both normoxia and hypoxia. Evidently, hPSCs lack many transcriptomic hallmarks of naive and formative pluripotency (a phase between naive and primed states). Integrative transcriptome analysis suggests our primed and RSeT hESCs are close to the early stage of post-implantation embryos, similar to the previously reported primary hESCs and early hESC cultures. Moreover, RSeT hESCs did not express naive surface markers such as CD75, SUSD2, and CD130 at a significant level. Biochemically, RSeT hESCs exhibit a differential dependency of FGF2 and co-independency of both Janus kinase (JAK) and TGFbeta signaling in a cell-line-specific manner. Thus, RSeT hESCs represent a previously unrecognized pluripotent state downstream of formative pluripotency. Our data suggest that human naive pluripotent potentials may be restricted in RSeT medium. Hence, this study provides new insights into pluripotent state transitions in vitro.

中文翻译：

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RSeT 人胚胎干细胞对幼稚和形成性多能性转化的抵抗力

人类胚胎干细胞 (hESC) 最重要的特性之一与其启动和初始多能状态有关。我们之前的荟萃分析表明存在源自不同朴素协议的异质多能状态。在这项研究中，我们描述了在各种生长条件下基于商业培养基（RSeT）的多能状态。值得注意的是，RSeT hESC 可以规避初始 hESC 所需的低氧生长条件，其中一些 RSeT 细胞（例如 H1 细胞）表现出低得多的单细胞铺板效率，在常氧和缺氧下都改变或大大延迟了细胞生长。显然，hPSC 缺乏许多幼稚和形成多能性（幼稚状态和启动状态之间的阶段）的转录组学特征。综合转录组分析表明，我们的引发和 RSeT hESC 接近植入后胚胎的早期阶段，类似于之前报道的原代 hESC 和早期 hESC 培养物。此外，RSeT hESC 不表达显着水平的初始表面标记，例如 CD75、SUSD2 和 CD130。从生化角度来看，RSeT hESCs 以细胞系特异性的方式表现出对 FGF2 的差异依赖性以及 Janus 激酶 (JAK) 和 TGFbeta 信号传导的共同独立性。因此，RSeT hESC 代表了形成性多能性下游的先前未被识别的多能状态。我们的数据表明，人类原始多能潜能可能在 RSeT 培养基中受到限制。因此，这项研究为体外多能状态转变提供了新的见解。