Synthetic Antifreeze Glycoproteins for Cellular Cryopreservation

Non-technical Abstract: Animals living in polar regions have adapted to survive freezing conditions. One of their strategies is to produce specialized antifreeze proteins that stop ice from forming in their blood and cells. These proteins prevent damage and death caused by ice crystals. When tissues and cells are frozen for medical and research purposes, chemicals are added to prevent ice from forming and causing damage. However, these chemicals can be harmful to cells that are important for medical purposes, such as stem cells, and there is a need for better strategies. The goal of this research project is to create polymer materials that work like the antifreeze proteins found in animals. The materials will be tested to see if they can stop ice crystals from forming and whether cells can survive being frozen in the presence of these materials. We will also examine if there are any toxic effects or genetic changes. As part of this research project, undergraduate and graduate students will be trained and all investigators will participate in community outreach efforts. This will include support and training efforts to females and underrepresented groups to encourage them to pursue careers in STEM. Technical Abstract: Extremophile organisms in cold climates produce specialized antifreeze glycoproteins (AFGPs) to defend their tissues against freezing damage. These proteins have shown great potential for prevention of cryopreservation damage of cells and tissues with research and regenerative medicine applications. However, AFGPs are heterogeneous in structure and can only be isolated in small quantities from the blood of polar fish. The overall goals of this project are to generate transformative materials that capture the chemical and biophysical properties of natural AFGPs and probe their ice binding and cryopreservation activity. This will be achieved by developing a scalable and tunable synthetic route to sAFGP structural variants and studying their ice binding, ice shaping, and membrane stabilizing properties. We will also examine bioactivity, including cellular viability and function after cryopreservation, cellular localization, any genetic changes, and induction of metabolic stasis. Additionally, we aim to incorporate EDI principles into biomaterials and glycoscience education and mentoring in the research lab, the classroom, and in community outreach. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.