Cryopreservation aims to revolutionize organ transplantation by enabling long-term storage of organs, thus overcoming geographical and time constraints for organ transplants. Vitrification, an alternative to traditional freezing, rapidly cools organs to a stable, ice-free, glass-like state, thus preventing osmotic damage. Cryoprotective agent (CPA) cytotoxicity prevents use of CPAs at concentrations needed to vitrify organs. This highlights the need for effective CPAs that form glass at non-cytotoxic concentrations. Our lab has developed a novel method which uses a 96-well plate and custom plate cooler to test the glass forming abilities of CPAs. This allows high throughput screening of potential CPAs to accelerate the search for good glass formers. Our plate cooler consisted of an aluminum block partially submerged in liquid nitrogen, and a copper plate on top of the aluminum. Each plate was placed on top of the copper to maximize contact with the bottom of the plate to the cooling system. Each well cooled at a consistent rate (8-15°C). We assessed each well’s status by eye: 0 for ice formation, 1 for glass formation, and categorized partially vitrified or cracked CPAs accordingly. Using this method, we tested 192 CPA solutions to determine if they’d vitrify. Our data aligned reasonably with similar works, suggesting this method can produce reliable and repeatable data. Future works will focus on automating data analysis and using acquired data to train a machine learning model to predict good glass formers.
Cameron Sugden is a PhD student in the Higgins lab at OSU. He previously graduated with a B.S. in molecular biology from Eastern Oregon University. His research focuses on complex biological systems and cryobiology. He employs methodologies from machine learning, dynamical systems, network theory, and computational biology, using a mix of data-driven and model-driven approaches.