Using metabolic pausing for maintaining stable and high-quality pluripotent stem cells

Regenerative medicine-based therapies have the potential to cure numerous currently untreatable using available regimens. Current therapies undergoing clinical trials include treatments for diseases ranging from diabetes to neurological disorders which primarily use pluripotent stem cells (PSCs). PSCs are a special and transient cell type that develops early in embryonic development and generates every other cell type in the body. It is now relatively easy to capture this stem cell state in vitro and PSCs are routinely established from humans and mice across laboratories. However, current culture conditions invariably lead to the generation of sub-optimal and unstable PSCs, making them inapt for either basic or translational studies.

Interestingly, several animals survive extreme environmental conditions by reducing their metabolic activity, body temperature, and respiration rate, processes known as diapause or hibernation. Here, we harness these evolutionarily conserved pathways for inducing a paused metabolic state using pharmacological tools, to allow effortless maintenance of good quality PSCs. We developed a novel culture condition that allowed metabolic pausing of PSCs in vitro. Further, we also extended this method for room temperature shipping of PSCs, thus eliminating the once indispensable cold-chain infrastructure required for their shipping. We also show the feasibility of our novel media condition in enabling the storage of mouse embryos for several days at room temperature. Such a method could, therefore, have broad application across cell therapy regimes requiring cell shipments, to fertility clinics, benefiting the lives of millions of patients.