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Related Experiment Videos

Trehalose expression confers desiccation tolerance on human cells.

N Guo1, I Puhlev, D R Brown

  • 1Center for Molecular Genetics, UCSD School of Medicine, La Jolla, CA 92093-0634, USA.

Nature Biotechnology
|February 5, 2000
PubMed
Summary

Human cells engineered to produce trehalose can survive dehydration. This trehalose production allows mammalian cells to retain viability in the dry state, offering potential for preserving biological materials.

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Area of Science:

  • Cell biology
  • Biochemistry
  • Biotechnology

Background:

  • Trehalose, a disaccharide, is known to confer desiccation tolerance in many organisms.
  • Understanding trehalose's protective mechanisms is crucial for cell preservation.
  • Previous research highlights trehalose's role in anhydrobiosis (life without water).

Purpose of the Study:

  • To investigate the feasibility of expressing trehalose biosynthetic genes in human cells.
  • To determine if trehalose production enhances the desiccation tolerance of human primary fibroblasts.
  • To assess the viability of engineered human cells after prolonged periods without water.

Main Methods:

  • Utilized a recombinant adenovirus vector for gene delivery of Escherichia coli otsA and otsB genes into human primary fibroblasts.

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  • Quantified trehalose production in infected cells, correlating it with the multiplicity of infection (MOI).
  • Assessed cell viability after desiccation using Fourier transform infrared spectroscopy to detect residual water content.

    • Main Results:

    • Successful expression of trehalose biosynthetic enzymes and increased trehalose accumulation in human fibroblasts.
    • Engineered cells exhibited enhanced survival, remaining viable for up to five days in a dry state.
    • Fourier transform infrared spectroscopy confirmed the absence of detectable water in dry, viable cells.

    Conclusions:

    • Mammalian cells can be successfully engineered to produce trehalose.
    • Trehalose expression confers significant desiccation tolerance to human primary fibroblasts.
    • This study demonstrates the potential for engineering mammalian cells to survive dehydration, with implications for cell preservation and regenerative medicine.