Expanding the cryoprotectant toolbox in biomedicine by multifunctional antifreeze peptides

The global cryopreservation market size rises exponentially due to increased demand for cell therapy-based products, assisted reproductive technology, and organ transplantation. Cryoprotectants (CPAs) are required to reduce ice-related damage, osmotic cell injury, and protein denaturation. Antioxidants are needed to hamper membrane lipid peroxidation under freezing stress, and antibiotics are added to the cryo-solutions to prevent contamination. The vitrification process for sized organs requires a high concentration of CPA, which is hardly achievable using conventional penetrating toxic CPAs like DMSO. Antifreeze peptides (AFpeps) are biocompatible CPAs leveraging inspiration from nature, such as freeze-tolerant and freeze-avoidant organisms, to circumvent logistic limitations in cryogenic conditions. This study aims to introduce the advances of AFpeps with cell-penetrating, antioxidant, and antimicrobial characteristics. We herein revisit the placement of AFpeps in the biobanking of cancer cells, immune cells, stem cells, blood cells, germ cells (sperms and oocytes), and probiotics. Implementing low-immunogenic AFpeps for allograft cryopreservation minimizes HLA mismatching risk after organ transplantation. Applying AFpeps to formulate bioinks with optimal rheology in extrusion-based 3D cryobiopriners expedites the bench-to-beside transition of bioprinted scaffolds. This study advocates that the fine-tuned synthetic or insect-derived AFpeps, forming round blunt-shape crystals, are biomedically broad-spectrum, and cell-permeable AFpeps from marine and plant sources, which result in sharp ice crystals, are appropriate for cryosurgery. Perspectives of the available room for developing peptide mimetics in favor of higher activity and stability and peptide-functionalized nanoparticles for enhanced delivery are delineated. Finally, antitumor immune activation by cryoimmunotherapy as an autologous in-vivo tumor lysate vaccine has been illustrated.