Establishment of Epidermal Cell Lines Derived from the Skin of the Atlantic Bottlenose Dolphin (Tursiops Truncatus).

Blake C. Ellis^1,2, Jin Yu², Mark S. Kindy^1,2,6, John E. Baatz^1,3, Margie Peden-Adams^1,3, Tara J. Ellingham⁴, Daynna J. Wolff⁴, Patricia A. Fair^1,7, and Sebastiano Gattoni-Celli^1,5,6*

(1)  Marine Biomedicine and Environmental Sciences Center, Charleston, SC

(2) Department of Neurosciences and Neuroscience Institute

(3) Department of Pediatrics

(4) Department of Pathology and Laboratory Medicine

(5) Department of Radiation Oncology, Medical University of South Carolina, Charleston, SC

(6) Ralph H. Johnson VA Medical Center, Charleston, SC

(7) NOAA/National Ocean Service, Center for Coastal Environmental Health and Biomolecular Research, Charleston, SC

The Atlantic bottlenose dolphin (BND) has become a focus of attention as an indicator of the environmental conditions of Atlantic coastal waters because of pathologies and diseases previously unseen in these marine mammals.  Humans are exposed to these same waters suggesting that dolphins may also be sentinels for human health, especially considering similarities in life span, offspring number, and growth and maturity rates between the two species.  Since the epidermis serves as the critical interface between the dolphin and its aquatic environment, we have established BND epidermal cell cultures and cell lines from skin tissue as an in vitro tool for evaluating environmental stressors on this protected marine mammal.  We have characterized the cell cultures by karyotype analysis, immunohistochemical staining for cytokeratin, and two-dimensional polyacrylamide gel electrophoresis, all of which revealed similar patterns between the cells and skin tissue.  Cell lines were obtained by transfection of the cell cultures with a plasmid encoding the SV40 small t and large T antigens in conjunction with the neomyocin-resistance gene.  Neomyocin-resistant clones exhibited a marked increase in growth rate compared to the non-transfected cell cultures.  We also found that dolphin cultures demonstrated greater tolerance to high salinity conditions compared to both the dolphin cell lines and the human keratinocyte cell line, HaCaT, as indicated by MTT assay.  The availability of BND epidermal cell cultures and cell lines provides a much-needed tool for comparing the responses of marine organisms and humans to environmental stressors as well as a novel experimental approach to studying dolphin skin.