Development of stabilizers for live cell preservation at room temperature (779.10)

Abstract

Cyropreservation at ultra‐low temperatures utilizing freezing mediums containing DMSO is the current best practice for long term storage of mammalian cells. Such conditions can impart devastating losses caused by common failures of the cold storage systems as well as the sensitivity of valuable cell types both primary and engineered to the freezing and thawing process. Hence recovery of viable cells from the frozen state is challenging and as is typical for many cell types only a fraction of the cells survive with the fully desired phenotype. Based on the studies of extremophiles such as tardigrades, rotifers or brine shrimp that can survive for many years in the dry state we hypothesized that techniques and protocols could be developed that mimic this molecular phenomena in eukaryotic cell cultures. Many previous candidate approaches or the last 15 years have failed to preserve cells under these conditions over the short term and no method to date has been successful at preserving cells under such conditions for more than a few hours. We applied a combinatorial screening approach to identify formulations containing both biostabilizers that preserve air‐dried eukaryotic cells at ambient temperature. The use specific biostabilizers in specific formulations protect cells from the stress of dehydration, dry‐storage, and rehydration while preventing the execution of apoptosis programs. We report here that we are able to achieve statistically significant improvements in cell survival rates and preservation time as compared to conventionally published dry down methods. These results provide a roadmap for achieving room temperature storage of mammalian cells in a dry state. The technology can be further developed for products that extend cell survival at ambient temperatures. This technology could eventually be applied to ambient storage of eukaryotic cell stocks for biomedical research, toxicology screens, cell‐based therapeutics and blood supply logistics.