Dry state preservation of nucleated cells: progress and challenges.

Effective stabilization of nucleated cells for dry storage would be a transformative development in the field of cell-based biosensors and biotechnologic devices, as well as regenerative medicine and other areas in which stem cells have clinical utility. Ultimately, the tremendous promise of cell-based products will only be fully realized when stable long-term storage becomes available without the use of liquid nitrogen and bulky, energetically expensive freezers. Significant progress has been made over the last 10 years toward this goal, but obstacles still remain. Loading cells with the protective disaccharide trehalose has been achieved by several different techniques and has been shown to increase cell survival at low water contents. Likewise, the protective effect of heat shock proteins and other compounds have also been explored alone and in combination with trehalose. In some cases, the benefit of these molecules is seen not initially upon rehydration, but over time during cellular recovery. Other considerations, such as inhibiting apoptosis and utilizing isotonic buffer conditions have also provided stepwise increases in cell viability and function following drying and rehydration. In all these cases, however, a low level of residual water is required to achieve viability after rehydration. The most significant remaining challenge is to protect nucleated cells such that this residual water can be safely removed, thus allowing vitrification of intra- and extracellular trehalose and stable dry state storage at room temperature.