Marine Biology Institute, College of Science Shantou University, Shantou, Guangdong, P.R. China.
Faculty of Veterinary and Animal Sciences, Lasbela University of Agriculture, Water and Marine Sciences, Uthal, Balochistan, Pakistan.
PLoS One. 2018 Mar 29;13(3):e0195128. doi: 10.1371/journal.pone.0195128. eCollection 2018.
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.
侏虎鲸（Feresa attenuata）（PKW）是一种热带和亚热带海洋哺乳动物，常见于大西洋、印度洋和太平洋。由于 PKW 生活在近海保护区，它们很少在陆地上被看到。因此，PKW 是最不为人知的海洋齿鲸物种之一。皮肤组织主要来自搁浅事件，这些事件发生在中国广东汕头沿海。采集的组织立即进行处理，并附着在涂有胶原蛋白的 6 孔组织培养板上。将完全培养基（DMEM 和 Ham's F12、胎牛血清、抗生素和必需氨基酸）添加到培养板中。原代培养（PKW-LWH）细胞通过波形蛋白和核型分析被验证为成纤维细胞，核型分析显示 42 条常染色体和两条性染色体 X 和 Y。将编码 SV40 大 T 抗原的质粒转染 PKW-LWH 细胞后，分离并扩增转染细胞。通过 RT-PCR、western blot、免疫荧光分析和 SV40 大 T 抗原稳定性的检测，证实了这一点。成纤维细胞 PKW-LWHT 的细胞增殖率比原代细胞 PKW-LWH 快，倍增时间分别为 68.9h 和 14.4h。在这项研究中，我们首次建立了侏虎鲸皮肤成纤维细胞系，为研究环境污染物和病原体对 PKW 和/或鲸类的影响提供了一个独特的体外研究机会。
