Format

Send to

Choose Destination
Cell Biol Toxicol. 2019 Jan 10. doi: 10.1007/s10565-018-09457-1. [Epub ahead of print]

Lifetime extension of humpback whale skin fibroblasts and their response to lipopolysaccharide (LPS) and a mixture of polychlorinated biphenyls (Aroclor).

Author information

1
School of Environment and Science, Griffith University, Brisbane, QLD, Australia.
2
Swiss Federal Institute of Aquatic Science and Technology (Eawag), Überlandstrasse 133, CH-8600, Dübendorf, Switzerland.
3
School of Veterinary Science, The University of Queensland, Gatton, QLD, Australia.
4
Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St Lucia, QLD, Australia.
5
Swiss Federal Institute of Aquatic Science and Technology (Eawag), Überlandstrasse 133, CH-8600, Dübendorf, Switzerland. Kristin.Schirmer@eawag.ch.
6
Institute of Biogechemistry and Pollutant Dynamics, ETH Zürich, Zürich, Switzerland. Kristin.Schirmer@eawag.ch.
7
School of Architecture, Civil and Environmental Engineering, EPF Lausanne, Lausanne, Switzerland. Kristin.Schirmer@eawag.ch.

Abstract

Marine mammals, such as whales, have a high proportion of body fat and so are susceptible to the accumulation, and associated detrimental health effects, of lipophilic environmental contaminants. Recently, we developed a wild-type cell line from humpback whale fibroblasts (HuWa). Extensive molecular assessments with mammalian wild-type cells are typically constrained by a finite life span, with cells eventually becoming senescent. Thus, the present work explored the possibility of preventing senescence in the HuWa cell line by transfection with plasmids encoding the simian virus large T antigen (SV40T) or telomerase reverse transcriptase (TERT). No stable expression was achieved upon SV40 transfection. Transfection with TERT, on the other hand, activated the expression of telomerase in HuWa cells. At the time of manuscript preparation, the transfected HuWa cells (HuWaTERT) have been stable for at least 59 passages post-transfection. HuWaTERT proliferate rapidly and maintain initial cell characteristics, such as morphology and chromosomal stability. The response of HuWaTERT cells to an immune stimulant (lipopolysaccharide (LPS)) and an immunotoxicant (Aroclor1254) was assessed by measurement of intracellular levels of the pro-inflammatory cytokines interleukin (IL)-6, IL-1β and tumour necrosis factor (TNF)-α. HuWaTERT cells constitutively express IL-6, IL-1β and TNFα. Exposure to neither LPS nor Aroclor1254 had an effect on the levels of these cytokines. Overall, this work supports the diverse applicability of HuWa cell lines in that they display reliable long-term preservation, susceptibility to exogenous gene transfer and enable the study of humpback whale-specific cellular response mechanisms.

KEYWORDS:

Cell line transfection; Humpback whale; Immunotoxicity; Inflammatory cytokines; Megaptera novaeangliae; Relative telomerase activity

PMID:
30627956
DOI:
10.1007/s10565-018-09457-1

Supplemental Content

Full text links

Icon for Springer
Loading ...
Support Center