Abstract Red blood cells (RBCs) are possibly the most transfused blood component and the most widely stored cell type. While cold storage (+4 oC) of RBCs has been vastly improved in the last few decades with a standard storage time of 42 days in clinical settings, recent clinical retrospective as well as laboratory studies indicate that beyond 14 days of storage RBCs might have vastly different biochemical properties and possibly inferior outcomes in patients. Here, we propose to develop an RBC preservation method based on our recent breakthrough in deep supercooling (DSC) of aqueous solutions where we can achieve seemingly stable supercooling for large volumes and at very low temperatures (down to -20 oC) for very long times. This is achieved by surface sealing of aqueous solutions via water-immiscible hydrocarbon-based liquids. Our central hypothesis, based on preliminary studies and prior work, is that DSC of RBCs can provide an immediately practical, high quality, and long-term storage, as an alternative to current clinical standard of cold storage. Our approach is to first establish a robust characterization framework for storage related injuries to a) establish the basic temperature optimization of DSC for RBCs, b) while comparing the cell quality of this basic approach to cold storage and cryopreservation. We will then supplement the DSC approach drawing from our experience in alleviating lipid peroxidation, oxidative stress, and membrane injuries and metabolic suppression along with recent advances in RBC preservation such as anaerobic preservation to achieve long-term storage of RBCs in small volumes. Finally, we aim to extend the range of parameters for DSC to the 150-500 ml range (volume), ~- 25-30 oC (temperature), and 150 days (time) in parallel. Our final goal is to conduct storage with these robust DSC strategies to preserve a clinical unit (~300 ml) of RBCs for 150 days. By completing this project, we expect, to demonstrate a novel method to dramatically extend the storage time for RBCs to 150 days whereby alleviating problems of current approaches. In the long-term the “large volume DSC” method, is widely applicable to all cell, tissue and organs; especially those that are not amenable to cryopreservation. These advances will positively influence healthcare by enabling storage of living matter for applications in cell/organ transplantation, engineered tissue logistics, and food storage among others

NARRATIVE: Red blood cells (RBCs) are possibly the most transfused blood component and the most widely stored cell type. While cold storage (+4 oC) of RBCs has been vastly improved in the last few decades with a standard storage time of 42 days in clinical settings, recent clinical retrospective as well as laboratory studies indicate that beyond 14 days of storage RBCs might have vastly different biochemical properties and possibly inferior outcomes in patients. Here, we propose to develop an RBC preservation method based on our recent breakthrough in deep supercooling of aqueous solutions where we can achieve seemingly stable supercooling for large volumes and at very low temperatures (down to -20 oC) for very long times. Our eventual goal, by combining our deep supercooling approach with other advances in RBC storage, is to extend the acceptable storage period to a minimum of 100 days to relieve logistical constraints, reduce waste of donated blood, and alleviate concerns related to use of aged cold-stored RBCs.