Oscillating Magnetic Field Assisted Supercooling Preservation of Fresh and Unfrozen Ovaries at Subzero Temperatures

项目总结/摘要 当材料在冻结过程中的内部温度低于其平衡冻结时 在冰成核发生之前的时间点，材料被称为处于过冷状态。的 本项目的长期目标是开发一种新型的商业上可行的过冷装置 将生物材料保存在远低于冰点的温度下， 功能活。振荡模式下的工程磁场已被证明 利用水的抗磁性控制水分子的放电和重新排列 分子。因此，生物材料中的水在零度以下的温度下可以保持不冻结 当施加磁场时。在许多其他用于冷冻保存和器官移植的材料中， 为了进行概念验证，我们选择了整个小鼠卵巢。的 开发的过冷技术将延长储存时间， 与经典冷冻保存相比，卵巢功能恢复率为零毒性 方法.在I期研究中， 过冷技术将被测试为一个扩展 小鼠卵巢在-5至-10 ℃的过冷状态 4周，并保存 将检查肥料。我们期待一个强大的和稳定的解决方案， 非冷冻保存各种生物样品，即细胞、组织和器官， 扩展名.在过去的几年里，一些研究已经出现了质疑的功能 使用常规低温保存方法储存的生物材料的质量。他们 表明（i）生物系统具有高度有组织的结构， 冷冻/解冻过程和（ii）高浓度的冷冻保护剂（CPA）， 因为二甲基亚砜（DMSO）具有潜在的毒性。当生物样品冷却到低于 细胞结构内的水的熔点或平衡冻结温度将 经历从液体到固体的相变。细胞外冰的形成被认为是 对结构化组织和器官的危害。此外，DMSO显示出高细胞毒性， 神经元样细胞、心肌细胞和粒细胞的分化，并且需要 解冻后迅速消除。所开发的技术将提供技术上的可行性， 任何完整器官或复杂组织保存工作的坚实基础， 潜在的结构和功能的影响，保存过程中的高含量，复杂的， 和人体器官。

PROJECT SUMMARY/ABSTRACT When the internal temperature of a material during freezing is below its equilibrium freezing point before ice nucleation has occurred, the material is said to be in the supercooled state. The long-term objective of this project is to develop a novel commercially viable supercooling device to preserve biological materials far below their freezing points while retaining their functionalities live. Engineered magnetic fields in the oscillation mode has been proved to control the discharge and realignment of water molecules using diamagnetic properties of water molecules. Therefore, water in biological materials can remain unfrozen at subzero temperatures when magnetic fields are applied. Among many other materials for cryopreservation and organ banking, we have selected the whole mice ovary for our focused proof of concept. The developed supercooling technology will enable an extended storage duration as well as higher recovery rates of ovarian functionalities with zero toxicity, compared to classic cryopreservation methods. In the Phase I study, the supercooling technology will be tested for an extended supercooling status of mice ovaries at -5 to -10 °C for up to 4 weeks, and their preserved fertilities will be examined. We anticipate a robust and stabilized solution for hypothermic yet non-freezing preservation of diverse biological samples, i.e. cells, tissues, and organs, by extension. In the past several years, a number of studies have appeared questioning the functional qualities of biological materials stored using conventional cryopreservation methods. They indicated that (i) biological systems have highly organized structures that are extremely sensitive to freezing/thawing processes and (ii) high concentrations of cryoprotective agents (CPAs) such as dimethyl sulfoxide (DMSO) are potentially toxic. When the biological sample is cooled below its melting point, or equilibrium freezing temperature, water within the cellular structure will undergo a phase change from liquid to solid. The formation of extracellular ice is known to be a hazard to structured tissues and organs. In addition, DMSO shows high cytotoxicity and affects the differentiation of neuron-like cells, cardiac myocytes, and granulocytes, and needs to be eliminated rapidly after thawing. The developed technique will offer the technical feasibility and solid foundations for any full-organ or complex tissue preservation efforts, providing insight into potential structural and functional effects of the preservation process on high-content, complex and human-derived organs.