A versatile polypharmacology platform promotes cytoprotection and viability of human pluripotent and differentiated cells

• DOI: 10.1038/s41592-021-01126-2
• Publication Date: 2021-05-03T16:03:40Z
• Authors: Yu Chen; Carlos A. Tristan; Lu Chen; Vukasin M. Jovanovic; Claire Malley; Pei-Hsuan Chu; Seungmi Ryu; Tao Deng; Pinar Ormanoglu; Dingyin Tao; Yuhong Fang; Jaroslav Slamecka; Hyenjong Hong; Christopher A. Le...; Sam Michael; Christopher P. Au...; Anton Simeonov; Ilyas Singeç
• Keywords: Cryopreservation; Pluripotent Stem Cells; 0301 basic medicine; rho-Associated Kinases; Cell Survival; Polypharmacology; Cell Culture Techniques; Cell Differentiation; Article; High-Throughput Screening Assays; 3. Good health; 03 medical and health sciences; Cryoprotective Agents; Gene Expression Regulation; Humans

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.