A small stress protein acts synergistically with trehalose to confer desiccation tolerance on mammalian cells

• Published in: Cryobiology Vol. 51; no. 1; pp. 15 - 28
• Main Authors: Ma, Xiaocui, Jamil, Kamran, MacRae, Thomas H., Clegg, James S., Russell, Joseph M., Villeneuve, Tania S., Euloth, Michelle, Sun, Yu, Crowe, John H., Tablin, Fern, Oliver, Ann E.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier Inc 01-08-2005
• Subjects: 293H cells; Air; Animals; Artemia; Blotting, Western; Cell culture; Cell Line; Cell Survival; Cryopreservation - methods; Dehydration; Desiccation; Desiccation tolerance; DNA, Complementary - metabolism; Dose-Response Relationship, Drug; Freeze Drying; Heat-shock proteins; Heat-Shock Proteins - metabolism; Humans; Microscopy, Fluorescence; Molecular Chaperones - metabolism; p26; Protein Denaturation; Time Factors; Transfection; Trehalose; Trehalose - chemistry; Water - chemistry; Heat-shock proteins; Cell culture; 293H cells; Trehalose; p26; Desiccation tolerance; Dehydration
• ISSN: 0011-2240; 1090-2392
• DOI: 10.1016/j.cryobiol.2005.04.007

The ability to desiccate mammalian cells while maintaining a high degree of viability would be very important in many areas of biological science, including tissue engineering, cell transplantation, and biosensor technologies. Certain proteins and sugars found in animals capable of surviving desiccation might aid this process. We report here that human embryonic kidney (293H) cells transfected with the gene for the stress protein p26 from
Artemia and loaded with trehalose showed a sharp increase in survival during air-drying. Further, we find vacuum-drying greatly improved the ability of the cells to survive, and that the physical shape and structure of the cellular sample had a large influence on recovery following rehydration. Cells suspended in a rounded droplet survived desiccation markedly better than those spread as a thin film. Finally, we used alamarBlue to monitor cellular metabolism and Hema 3 to assess colony formation after vacuum-drying. AlamarBlue fluorescence indicated that the transfected 293H cells expressing p26 (E11′L) grew much better than the control 293H cells. In fact, immediate survival and colony formation in E11′L cells increased as much as 34-fold compared with control cells when the samples were dried to a water content of 0.2 g H
2O/g dry weight, as measured by gravimetric analysis. These results indicate that p26 improves cell survival following drying and rehydration, and suggest that dry storage of mammalian cells is a likely possibility in the future.