Abstract Cryopreservation by vitrification typically requires 30–50 v/v% of cytotoxic penetrable cryoprotective agents (CPAs) to prevent ice crystal formation during freezing and thawing, limiting its broader application. Since pressure suppresses ice crystallization, applying high pressure during vitrification may enable reducing CPA concentrations while maintaining cell viability. In this study, we used a high pressure freezing (HPF) device, commonly used for cryofixation, to successfully cryo