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

摘要 红细胞（RBC）可能是输血最多的血液成分和储存最广泛的细胞 类型.虽然红细胞的冷藏（+4 ℃）在过去几十年中得到了极大的改善， 在临床环境中的储存时间为42天，最近的临床回顾性研究以及实验室研究表明， 储存超过14天的红细胞可能具有非常不同的生化特性， 患者的结果。在这里，我们建议开发一种红细胞保存方法，根据我们最近的 水溶液深度过冷（DSC）的突破，我们可以实现看似稳定的 过冷，适用于大体积和极低温度（低至-20 ° C）的长时间使用。这是 通过经由水不混溶的烃基液体的水溶液的表面密封来实现。我们的中央 基于初步研究和先前工作的假设是，RBC的DSC可以立即提供 实用，高质量和长期储存，作为当前临床标准冷藏的替代品。我们 一种方法是首先建立一个强大的表征框架，用于存储相关伤害，以a）建立 用于RBC的DSC的基本温度优化，B）同时将该基本方法的细胞质量与 冷藏和冷冻保存。然后，我们将根据我们在以下方面的经验，补充DSC方法： 减轻脂质过氧化、氧化应激和膜损伤以及代谢抑制，沿着 红细胞保存的最新进展，如厌氧保存，以实现红细胞的长期储存， 小体积。最后，我们的目标是将DSC的参数范围扩展到150-500 ml范围（体积）， 25-30 ℃（温度），150天（时间）平行。我们的最终目标是利用这些强大的 将临床单位（约300 ml）RBC保存150天的DSC策略。通过完成这个项目，我们预计， 证明了一种新的方法，可以将RBC的储存时间大大延长到150天， 目前的方法问题。在长期的“大体积DSC”方法中，广泛适用于所有的电池， 组织和器官;特别是那些不适合冷冻保存的组织和器官。这些进步将积极 通过能够储存用于细胞/器官移植的生物物质来影响医疗保健， 组织物流和食物储存等等

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