Protection of CANARY Cells After Drying and Rehydration Correlates with Decrease in Apoptotic Cell Death

Abstract

CANARY cells are genetically engineered murine B cells that serve as a rapid detection system for various infectious pathogens. We are attempting to produce a stable dehydrated cellular product to make this system practical. A major source of damage to these cells during drying was identified as apoptosis. Trehalose, which protects mammalian cells during drying, was investigated with regard to its effect on apoptosis. B cells were loaded with trehalose and vacuum-dried. Trehalose reduced apoptosis during drying and rehydration, and when used in combination with a pan-caspase inhibitor OPH-109, the degree of apoptotic cell loss was further diminished. In this case, viability following rehydration reached 70%–80%. Surprisingly, trehalose alone blocked apoptotic cell death better than OPH-109 alone (45% versus 70% total apoptotic cells for trehalose- or OPH-109–treated samples, respectively, after drying to 0.3 g H2O/g dry weight). Nevertheless, optimal viability was achieved when the cells were loaded and d...