Insights on cryoprotectant toxicity from gene expression profiling of endothelial cells exposed to ethylene glycol

Cryobiology. 2015 Dec;71(3):405-12. doi: 10.1016/j.cryobiol.2015.10.142. Epub 2015 Oct 22.


Cryopreservation consists of preserving living cells or tissues generally at -80 °C or below and has many current applications in cell and tissue banking, and future potential for organ banking. Cryoprotective agents such as ethylene glycol (EG) are required for successful cryopreservation of most living systems, but have toxic side effects whose mechanisms remain largely unknown. In this work, we investigated the mechanisms of toxicity of ethylene glycol in human umbilical vein endothelial cells (HUVECs) as a model of the vascular endothelium in perfused organs. Exposing cells to 60% v/v EG for 2 h at 4 °C resulted in only a slight decrease in subsequent cell growth, suggesting only modest toxicity of EG for this cell type. Gene expression analysis with whole genome microarrays revealed signatures indicative of a generalized stress response at 24 h after EG exposure and a trend toward partial recovery at 72 h. The observed changes involved signalling pathways, glycoproteins, and genes involved in extracellular and transmembrane functions, the latter suggesting potential effects of ethylene glycol on membranes. These results continue to develop a new paradigm for understanding cryoprotectant toxicity and reveal molecular signatures helpful for future experiments in more completely elucidating the toxic effects of ethylene glycol in vascular endothelial cells and other cell types.

Keywords: Antifreeze; Cell model; Cryobiology; Cryopreservation; Functional genomics; High throughput; Toxicology.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cryopreservation / methods*
  • Cryoprotective Agents / toxicity*
  • Endothelial Cells / drug effects*
  • Ethylene Glycol / toxicity*
  • Gene Expression Profiling
  • Humans
  • Microarray Analysis
  • Transcriptome / drug effects*


  • Cryoprotective Agents
  • Ethylene Glycol