Abstract
The pygmy killer whale (Feresa attenuata) (PKW) is a tropical and subtropical marine mammal commonly found in the Atlantic, Indian and Pacific oceans. Since the PKWs live in offshore protected territories, they are rarely seen onshore. Hence, PKW are one of the most poorly understood oceanic species of odontocetes. The dermal tissue comes primarily from stranding events that occur along the coast of the Shantou, Guangdong, China. The sampled tissues were immediately processed and attached on collagen-coated 6-well tissue culture plate. The complete medium (DMEM and Ham’s F12, fetal bovine serum, antibiotic and essential amino acids) was added to the culture plates. The primary culture (PKW-LWH) cells were verified as fibroblast by vimentin and karyotype analyses, which revealed 42 autosomes and two sex chromosomes X and Y. Following transfection of PKW-LWH cells with a plasmid encoding, the SV40 large T-antigens and the transfected cells were isolated and expanded. Using RT-PCR, western blot, immunofluorescence analysis and SV40 large T-antigen stability was confirmed. The cell proliferation rate of the fibroblast cells, PKW-LWHT was faster than the primary cells PKW-LWH with the doubling time 68.9h and 14.4h, respectively. In this study, we established PKW dermal fibroblast cell line for the first time, providing a unique opportunity for in vitro studies on the effects of environmental pollutants and pathogens that could be determined in PKW and/or Cetaceans.
Highlights
Pygmy killer whales (Feresa attenuata) are tropical and subtropical delphinid with extremely aggressive behaviour, they are rarely seen onshore and are one of the most poorly known oceanic species of odontocetes [1]
Considering the critical importance of cell culturing and genetic material preservation in conservation biology laboratories, we focused on establishing a PKW cell line, which will help in broadening research strategies and offer researchers a reliable tool for understanding the biological response and mechanisms of PKW and/or other marine mammals
On the 22nd day of primary culturing, morphologically fibroblastic cells with polygonal and spindle-like shaped appeared in the culture plates and were photographed under phase contrast microscope (Fig 1)
Summary
Pygmy killer whales (Feresa attenuata) are tropical and subtropical delphinid with extremely aggressive behaviour, they are rarely seen onshore and are one of the most poorly known oceanic species of odontocetes [1]. PKW habitats in deep and warm waters beyond the onshore continent, and very rarely seen close to the edges of the continental shelf (except near few island, where the water is very deep). Current population estimation of PKW by IUCN, approximately 38,900, is for the eastern tropical Pacific population. [3], whereas in the eastern. Current population estimation of PKW by IUCN, approximately 38,900, is for the eastern tropical Pacific population. [3], whereas in the eastern
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