title: Insights on cryoprotectant toxicity from gene expression profiling of endothelial cells exposed to ethylene glycol.
abstract: 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.
authors.affiliation: Integrative Genomics of Ageing Group, Institute of Integrative Biology, University of Liverpool, Liverpool L69 7ZB, UK.
authors.forename: Rui Martins
authors.initials: RM
authors.lastname: Cordeiro
authors.affiliation: Integrative Genomics of Ageing Group, Institute of Integrative Biology, University of Liverpool, Liverpool L69 7ZB, UK.
authors.forename: Soren
authors.initials: S
authors.lastname: Stirling
authors.affiliation: 21st Century Medicine, Inc., Fontana, CA 92336, USA.
authors.forename: Gregory M
authors.initials: GM
authors.lastname: Fahy
authors.affiliation: Integrative Genomics of Ageing Group, Institute of Integrative Biology, University of Liverpool, Liverpool L69 7ZB, UK. Electronic address: jp@senescence.info.
authors.forename: João Pedro
authors.initials: JP
authors.lastname: de Magalhães
mesh_terms: D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000633:toxicity*; D042783:Endothelial Cells / Q000187:drug effects*; D019855:Ethylene Glycol / Q000633:toxicity*; D020869:Gene Expression Profiling; D006801:Humans; D046228:Microarray Analysis; D059467:Transcriptome / Q000187:drug effects*
chemical_list: D003451:Cryoprotective Agents; D019855:Ethylene Glycol
keywords: Antifreeze; Cell model; Cryobiology; Cryopreservation; Functional genomics; High throughput; Toxicology