Human pluripotent stem cells (hPSCs) are capable of extensive self-renewal yet remain highly sensitive to environmental perturbations in vitro, posing challenges to their therapeutic use. There is an urgent need to advance strategies that ensure safe and robust long-term growth and functional differentiation of these cells. Here, we deployed high-throughput screening strategies to identify a small-molecule cocktail that improves viability of hPSCs and their differentiated progeny. The combination of chroman 1, emricasan, polyamines, and trans-ISRIB (CEPT) enhanced cell survival of genetically stable hPSCs by simultaneously blocking several stress mechanisms that otherwise compromise cell structure and function. CEPT provided strong improvements for several key applications in stem-cell research, including routine cell passaging, cryopreservation of pluripotent and differentiated cells, embryoid body (EB) and organoid formation, single-cell cloning, and genome editing. Thus, CEPT represents a unique poly-pharmacological strategy for comprehensive cytoprotection, providing a rationale for efficient and safe utilization of hPSCs.

人类多能干细胞 (hPSCs) 能够进行广泛的自我更新，但对体外环境扰动仍然高度敏感，这对其治疗用途提出了挑战。迫切需要推进确保这些细胞安全和稳健的长期生长和功能分化的策略。在这里，我们部署了高通量筛选策略来确定一种小分子混合物，该混合物可提高 hPSC 及其分化后代的活力。chroman 1、emricasan、多胺和 trans-ISRIB (CEPT) 的组合通过同时阻断几种会损害细胞结构和功能的应激机制来增强遗传稳定的 hPSC 的细胞存活。CEPT 为干细胞研究中的几个关键应用提供了强大的改进，包括常规细胞传代，多能和分化细胞的冷冻保存、胚状体 (EB) 和类器官的形成、单细胞克隆和基因组编辑。因此，CEPT 代表了一种独特的综合细胞保护的多药理学策略，为高效和安全地利用 hPSC 提供了理论基础。