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

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

Nature methods (2021-05-05)
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 LeClair, Sam Michael, Christopher P Austin, Anton Simeonov, Ilyas Singeç
ABSTRACT

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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Donkey serum
Sigma-Aldrich
Chroman 1, ≥98% (HPLC)
Sigma-Aldrich
Anti-Puromycin Antibody, clone 12D10, clone 12D10, from mouse
Sigma-Aldrich
Antioxidant Supplement (1000×), liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Polyamine Supplement (1000×), liquid, sterile-filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
Anti-Myosin-10/MYH10/Myosin IIB Antibody, from rabbit, purified by affinity chromatography
Sigma-Aldrich
Anti-Sox2 Antibody, Chemicon®, from rabbit