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

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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.