Deep Supercooling of Red Blood Cells: Towards Practical Long Term Storage

摘要 红细胞(RBC)可能是输血最多的成分，也是储存最广泛的细胞 打字。虽然红细胞的冷藏(+4oC)在过去几十年里有了很大的改善，但标准 在临床环境中储存42天，最近的临床回顾以及实验室研究表明 超过14天的储存后，红细胞的生化特性可能会有很大的不同，甚至可能是劣质的 患者的结局。在这里，我们建议开发一种基于我们最近的RBC保存方法 在水溶液深度过冷(DSC)方面的突破，我们可以实现看似稳定的 过冷，适用于大容量和极低温度(低至-20摄氏度)很长时间。这是 通过不溶于水的碳氢化合物液体对水溶液进行表面密封。我们的中央 基于前期研究和前人工作的假设是，红细胞的DSC可以立即提供一个 实用、高品质、长期储存，作为目前临床冷藏标准的替代。我们的 方法是首先为与储存相关的伤害建立一个稳健的表征框架，以a)建立 红细胞差示扫描量热法的基本温度优化，b)同时比较这种基本方法的细胞质量 冷藏和超低温保存。然后，我们将从我们的经验中补充DSC方法 减轻脂质过氧化、氧化应激、膜损伤和代谢抑制 红细胞保存的最新进展，如厌氧保存以实现红细胞的长期保存 体积很小。最后，我们的目标是将DSC的参数范围扩大到150-500毫升(体积)，~- 25-30℃(温度)和150天(时间)平行。我们的最终目标是使用这些强大的 DSC策略将临床单位(~300毫升)的红细胞保存150天。通过完成这个项目，我们预计， 演示了一种新方法，将红细胞的存储时间大幅延长至150天，从而缓解了 当前方法的问题。从长远来看，“大容量DSC”方法广泛适用于所有单元格， 组织和器官；尤其是那些不适合冷冻保存的组织和器官。这些进展将是积极的 通过为细胞/器官移植应用实现生物物质存储来影响医疗保健 组织物流、食品储存等

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