Human extended pluripotent stem cells (EPSCs), with bidirectional chimeric ability to contribute to both embryonic and extraembryonic lineages, can be obtained and maintained by converting conventional pluripotent stem cells using chemicals. However, the transition system is based on inactivated mouse fibroblasts, and the underlying mechanism is not clear. Here we report a Matrigel-based feeder-free method to convert human embryonic stem cells and induced pluripotent stem cells into EPSCs and demonstrate the extended pluripotency in terms of molecular features, chimeric ability, and transcriptome. We further identify chemicals targeting glycolysis and histone methyltransferase to facilitate the conversion to and maintenance of feeder-free EPSCs. Altogether, our data not only establish a feeder-free system to generate human EPSCs, which should facilitate the mechanistic studies of extended pluripotency and further applications, but also provide additional insights into the transitions among different pluripotent states.

人类扩展多能干细胞 (EPSC) 具有双向嵌合能力以促进胚胎和胚胎外谱系，可以通过使用化学品转化常规多能干细胞来获得和维持。然而，过渡系统是基于灭活的小鼠成纤维细胞，其潜在机制尚不清楚。在这里，我们报告了一种基于基质胶的无饲养层方法，将人类胚胎干细胞和诱导多能干细胞转化为 EPSC，并在分子特征、嵌合能力和转录组方面证明了扩展的多能性。我们进一步确定了靶向糖酵解和组蛋白甲基转移酶的化学物质，以促进无饲养层 EPSC 的转化和维持。总而言之，我们的数据不仅建立了一个无饲养系统来生成人类 EPSC，