Grant: Project Grant

Preserving live cells outside the body for any extended period of time means that they must be stored at the temperature of liquid nitrogen, which is -196 °C. This low-temperature preservation is called cryopreservation, and it is necessary for the use of live cells in biological research for timing and cost. For cells to survive and be functional after the freezing and thawing processes, different cryopreservation methods must be specially developed and refined for each specific cell type. Cells frozen individually are used every day in research labs, transported by suppliers to end-users, or stored in cell banks for future transplantation. However, individual cells outside the body do not behave exactly the same way as cells inside the body where they are surrounded by, and interacting with, other cells. For this reason, multi-cell-type cultures are a much better outside-the-body biological model. Unfortunately, they are more difficult to cryopreserve than isolated cells. We are interested in developing ways to cryopreserve single-cell thick endothelial layers that line blood vessels, corneas, and intestines. Brain microvascular endothelial cells, together with astrocytes and pericytes, form the main functional component of the blood-brain barrier, which is the lining of blood vessels in the brain that allows nutrients in but keeps harmful organisms and most medications out. We will culture these three cell types together to make a blood-brain barrier analog and use it to develop new understanding of how ice forms inside cells during the cryopreservation of multi-cell-type 2D-layers. Using a systematic approach, we will develop methods to cryopreserve this and other blood-brain-barrier models. Cryopreservation of models of the blood-brain barrier will enable new research on drug delivery to the brain and on diseases associated with problems in the blood-brain barrier, such as Alzheimer's disease, Parkinson's disease, epilepsy, stroke, and multiple sclerosis.