Cell attachment defines sensitivity to cold stress via the Hippo pathway.

Although cells are frequently maintained at cold temperatures during experiments, the effects of cold stress on cell viability and subsequent cellular conditions remain elusive. In this study, we investigated the effects of cold stress on cancer cells under various culture conditions. We showed that cold stress induces ferroptosis, a form of cell death characterized by lipid peroxidation, in sensitive cancer cell lines. High cell density and serum starvation activate the Hippo pathway and suppress cold-induced cell death. Genetic deletion of Hippo pathway components enhances cold stress susceptibility. Furthermore, the cell attachment status influences the response to cold stress, with suspended cells showing greater resistance and faster recovery than attached cells. This study highlights the importance of cellular conditions and the Hippo pathway in the handling and storage of cancer cells at cold temperatures, thereby offering insights into experimental and clinical contexts.