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Trehalose expression confers desiccation tolerance on human cells

Abstract

Many organisms that withstand desiccation express the disaccharide trehalose. We have now expressed the otsA and otsB genes of Escherichia coli, which encode trehalose biosynthetic enzymes, in human primary fibroblasts using a recombinant adenovirus vector. Infected cells produced increased amounts of trehalose with increasing multiplicity of infection (MOI). Human primary fibroblasts expressing trehalose could be maintained in the dry state for up to five days. Fourier transform infrared spectroscopy indicated that dry, but viable, human cells contained no detectable water. This study shows that mammalian cells can be engineered to retain viability in the absence of water.

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Figure 1: HPLC analysis of trehalose.
Figure 2: (A) Trehalose production in 12F cells infected with Ad-OTS.
Figure 3: Photomicrograph of 12F cells dried for 24 h, rehydrated, and placed in CAM–EH1 solution to visualize live (green) and dead (orange) cells.
Figure 4: FTIR spectroscopy of hydrated and desiccated 12F cells.

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Acknowledgements

This research was supported by grants from the University of California BioSTAR program and Advanced Tissue Sciences Inc. (F.L.), and National Science Foundation Instrumentation and Laboratory Improvement Grant (D.B.). F.L. is a member of the University of California San Diego Cancer Center, the Whittier Institute for Diabetes, and the Department of Pediatrics. We gratefully acknowledge the assistance of Po Yee Leung, Enrique López, and Reneé Link with the FTIR assays.

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Correspondence to Fred Levine.

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Guo, N., Puhlev, I., Brown, D. et al. Trehalose expression confers desiccation tolerance on human cells. Nat Biotechnol 18, 168–171 (2000). https://doi.org/10.1038/72616

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