Cryopreservation in the Era of Cell Therapy: Revisiting Fundamental Concepts to Enable Future Technologies

Abstract Cryopreservation strives to maximize the viability and biofunctionality of cells and tissues by cooling them to a subzero temperature to facilitate storage and delivery. This technology has enabled clinics and labs to preserve rare and crucial samples and is poised to become more important with rising interest in cell therapy. Here, the biological impact of cooling rates on different cellular components is first described, paying special emphasis on the differences between slow cooling and vitrification with a heat transfer perspective based on the Biot number. This is followed by an overview of various classes of chemical‐based cryoprotective agents including small molecules, antifreeze proteins, hydrogels, and cryoprotective nanomaterials. Most importantly, fundamental concepts of cryopreservation including Mazur's “two‐factor hypothesis” are revisited, gaps in them are highlighted, and experiments to validate reported claims to deepen mechanistic understanding of cryoprotection are proposed. A matric is also introduced to assess the suitability of biomaterials for use in cell therapy to support manufacturers in making strategic choices for storing clinical samples. It is believed that this review would inspire readers to scrutinize fundamental concepts in cryopreservation to facilitate the development of new cryoprotective materials and technologies to support the emerging cell manufacturing and therapy industry.