Lessons from Nature: Antifreeze Glycolipids for Cryopreservation of Mammalian Cel

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DESCRIPTION (provided by applicant): Storage of cells for many researchers simply involves addition of 10% DMSO or similar cryoprotectants to cells in suspension, putting them in cryovials and slow cooling to subzero temperaturesf and storage in a mechanical or nitrogen cooled freezer or dewar. As long as viable cells are present upon thawing, cell yield may be a secondary consideration due to the high proliferative potential of the cells. However, there are significant cell types that are difficult to preserve with existing cryopreservation techniques including many primary cells, such as cardiac myocytes, and hepatocytes, plus embryonic stem cells and inducible pluripotent stem cells. Others, such as keratinocytes, are easy to cryopreserve in suspension but difficult when attached to a substrate. The study of how organisms survive extreme temperatures has revealed that multiple phenomena may occur in synchrony to promote survival including production of cryoprotectants, antifreeze compounds (peptides and lipids), dehydration and nucleators. Mammalian cell cryopreservation may be possible by mimicking strategies employed by certain insects to survive sub-zero temperatures. These insects combine increases in cryoprotectant content (such as glycerol) with production of antifreeze compounds, at the onset of cold environmental conditions, to survive temperatures as low as -80oC. The presence of these compounds lowers the freezing point of the solution and also changes the shape and formation of ice. In this way, potentially less cryoprotectants, such as DMSO or glycerol, may be used to preserve cells reducing potential cryoprotectant induced cytotoxicity concerns and offering alternative preservation methods for cells and tissues that can be used in clinical applications and cell and tissue-based screening assays for research applications.